ANRV253-MI59-06 ARI 4 August 2005 11:16

Alternative Candida albicans

Lifestyles: Growth on
Surfaces
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Carol A. Kumamoto and Marcelo D. Vinces

Department of Molecular Biology and Microbiology, Tufts University, Boston,
Massachusetts 02111; email: carol.kumamoto@tufts.edu, marcelo.vinces@tufts.edu

Annu. Rev. Microbiol. Key Words

2005. 59:113–33
biofilm, tissue invasion, invasive growth, thigmotropism, hyphae
First published online as a
Review in Advance on
May 2, 2005 Abstract
The Annual Review of Candida albicans, an opportunistic fungal pathogen, causes a wide
Microbiology is online at variety of human diseases such as oral thrush and disseminated can-
micro.annualreviews.org
didiasis. Many aspects of C. albicans physiology have been studied
doi: 10.1146/ during liquid growth, but in its natural environment, the gastroin-
annurev.micro.59.030804.121034
testinal tract of a mammalian host, the organism associates with
Copyright

c 2005 by surfaces. Growth on a surface triggers several behaviors, such as
Annual Reviews. All rights
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biofilm formation, invasion, and thigmotropism, that are important
for infection. Recent discoveries have identified factors that regulate
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0113$20.00
these behaviors and revealed the importance of these behaviors for
pathogenesis.

113
 ANRV253-MI59-06 ARI 4 August 2005 11:16

Contents
INTRODUCTION . . . . . . . . . . . . . . . . . 114 MECHANISMS OF INVASIVE
SURFACES COLONIZED BY GROWTH . . . . . . . . . . . . . . . . . . . . . . 119
CANDIDA ALBICANS . . . . . . . . . . . 115 Tissue Invasion by C. albicans
MECHANISMS OF BIOFILM Hyphae Occurs on Epithelial,
FORMATION AND Epidermal, and Endothelial
DEVELOPMENT. . . . . . . . . . . . . . . 115 Surfaces During
Biofilm Formation on Implanted Candidiasis. . . . . . . . . . . . . . . . . . . . 119
Medical Devices Results in Protease and Phospholipase
Drug Refractory Infections . . . . 115 Activities Contribute to
Numerous Parameters Influence Invasion of Host
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Biofilm Formation and Tissue . . . . . . . . . . . . . . . . . . . . . . . . 119

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Structure. . . . . . . . . . . . . . . . . . . . . . 116 Filamentation During Invasive

Multiple Mechanisms Contribute Growth is Promoted by
to the High Drug Resistance Physical Contact . . . . . . . . . . . . . . 120
of Biofilm Cells . . . . . . . . . . . . . . . 116 Transcription Factors Regulating
High Levels of Amino Acid Invasive Growth by Embedded
Biosynthesis and Protein Cells . . . . . . . . . . . . . . . . . . . . . . . . . . 120
Synthesis in Biofilm Signaling Pathways Regulating
Cells . . . . . . . . . . . . . . . . . . . . . . . . . . 117 Invasive Growth by Embedded
Defective Biofilm Development Cells . . . . . . . . . . . . . . . . . . . . . . . . . 123
Caused by Mutations that Models for Invasion of Tissue
Alter the Cell Surface and Surfaces . . . . . . . . . . . . . . . . . . . . . . . 124
Compromise Adherence . . . . . . . 118 GUIDANCE OF HYPHAE BY
Development of Animal Models THIGMOTROPISM . . . . . . . . . . . . 125
for Biofilms . . . . . . . . . . . . . . . . . . . 118 CONCLUSIONS . . . . . . . . . . . . . . . . . . . 126

INTRODUCTION perficial mucosal infections to invasive, life-

threatening disease. Candida spp. rank among
In the fourth century B.C., growth of
the four most common causes of bloodstream
Nosocomial: an the opportunistic fungal pathogen Candida
infection that infections and cardiovascular infections in
albicans on the surface of human tissue was
develops within a U.S. hospitals (18, 41). In neonatal intensive
noted and the oral infection it causes, thrush,
hospital and is care units, Candida spp. are an even more fre-
was described by Hippocrates. Since that
produced by an quent cause of bloodstream infections (95).
infectious organism time, the incidence of candidiasis has in-
The advances of modern medicine have led
acquired during the creased and C. albicans has become a sig-
stay of the patient to larger populations of compromised patients
nificant nosocomial pathogen. The modern
susceptible to candidiasis, increasing the im-
AIDS epidemic has created a population of
portance of C. albicans as a pathogen and
patients susceptible to candidiasis; oral thrush
providing impetus for the detailed study of
is one of the most common opportunistic in-
C. albicans biology.
fections in AIDS patients (4).
As is typical for many microorganisms,
As an opportunist, C. albicans does not
C. albicans physiology has been frequently
usually cause serious disease in immunocom-
studied during growth in liquid culture. How-
petent hosts, but immunodeficient hosts are
ever, in their natural environment, C. albicans
susceptible to infections ranging from su-

114 Kumamoto · Vinces

 ANRV253-MI59-06 ARI 4 August 2005 11:16
cells are commonly found in association
with surfaces. Therefore, it is important to
understand how C. albicans cells interact
with surfaces and how the unique aspects of
growth on surfaces contribute to C. albicans
pathogenicity. Several distinct C. albicans
behaviors occur on surfaces, including
biofilm formation, invasive growth, and
thigmotropism. In this review we describe
these behaviors and discuss the mechanisms
that regulate them. Each behavior occurs
under specific environmental conditions and
is mediated and controlled by specific fungal
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SURFACES COLONIZED BY
CANDIDA ALBICANS
C. albicans is generally found as a commen-
sal organism in association with a human or
animal host. In one study, 7.1% of infants
were colonized by Candida or other yeasts
on the day of birth and 96% of infants were
orally colonized by approximately one month
of age (94). In adults, gastrointestinal car-
riage of Candida spp. is common (78). For
example, fungi were found in 80% of fecal
samples from healthy adults (37). Soll et al.
(104) showed that several surfaces in the body
can be colonized, including the vaginal wall,
surfaces in the oral cavity, and the anorectal
surface.
Individuals carry a particular strain for
long periods, but changes in the colonizing
strain over time can also be detected (78). A
single individual may carry unrelated strains at
different sites (104), and changes in the colo-
nizing C. albicans strain have been observed in
HIV-positive patients (109). Thus, coloniza-
tion by C. albicans is not fixed and changes may
occur during an individual’s lifetime.
Within the host, a population of com-
mensal organisms is probably bound to mu-
cosal surfaces. In mice orally inoculated with
C. albicans, binding of yeast-form C. albicans
cells to epithelial surfaces in the gastrointesti-
nal tract was detected within 3 (82) or 72 h
(51) postinoculation. Another yeast species,
Torulopsis pintolopesii, naturally colonizes mice.
This organism is found in layers bound to
the secreting epithelium of the stomach (97).
Biofilm: a
These findings demonstrate the ability of community of
commensal fungal organisms to adhere to tis- microorganisms
sue surfaces within the host. attached to a surface,
In summary, growth on a biological sur- forming
three-dimensional
face, especially in the mammalian gastroin-
structures containing
testinal tract, is part of the natural lifestyle exopolymeric matrix
of C. albicans. We discuss below how the in- and cells that
teraction of C. albicans with a surface alters exhibit distinctive
C. albicans behavior and how these effects con- phenotypic
properties
tribute to disease.
Thigmotropism:
the directional
response of a cell or
MECHANISMS OF BIOFILM tissue to touch, or
FORMATION AND physical contact with
DEVELOPMENT a solid object
Epithelial:
Biofilm Formation on Implanted pertaining to tissue
Medical Devices Results in Drug that forms the outer
Refractory Infections surface of the body
and lines the body
A biofilm is a three-dimensional commu- cavities, and to main
nity of microorganisms embedded in an ex- tubes and
opolymeric matrix and attached to a surface. passageways that lead
Upon attachment, microorganisms undergo a to the exterior, such
as the lining of the
change to a sessile (attached) lifestyle. Dental
gastrointestinal tract
plaque is a well-known natural example of a
bacterial biofilm found in humans. C. albicans
also forms a biofilm on dental enamel (61) as
well as on human heart valves, causing endo-
carditis (29).
From a human health perspective, biofilms
are important because they form on implanted
medical devices and result in infections that
are unusually refractory to antimicrobial ther-
apy. Medical device infection contributes to
about half of all nosocomial infections (for
review see Reference 54). Several million
vascular and urinary catheters and tens of
thousands of prosthetic heart valves are used
annually in the United States; approximately
10% of infections linked to these devices are
due to Candida spp. (54). For example, of the
estimated 80,000 bloodstream infections asso-
ciated with central venous catheters that occur
annually in U.S. intensive care units, 11.5%
are due to Candida spp. (54). The mortality
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 ANRV253-MI59-06 ARI 4 August 2005 11:16
rate for device-associated Candida infection is
approximately 30% (54).
Cells in a biofilm characteristically exhibit
Quorum sensing:
cell-density- high resistance to antimicrobial drugs, and
dependent therefore infected devices must usually be re-
communication and moved to cure the infection (54). For some
coordination of devices, removal necessitates major surgery
microbial behavior
and exposes patients to significant risks. As a
via signaling
molecules result, the ability of C. albicans to adhere to
a medical device and form a biofilm resistant
to antifungal agents represents an important
medical challenge.
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Numerous Parameters Influence
Biofilm Formation and Structure
Studies have demonstrated that C. albicans
biofilms form in several stages (21, 29). First,
cells, typically yeast form, attach to a surface.
Second, cells proliferate on the surface, form-
ing microcolonies. Third, growth of cells,
production of hyphae (filamentous forms of
the organism), and secretion of exopolymeric
matrix result in elaboration of the characteris-
tic three-dimensional structure that is typical
of a biofilm. The exopolymeric matrix, com-
posed of carbohydrates, proteins, and other
unidentified components, surrounds the cells
in a mature biofilm (11).
Numerous materials and growth media
support the growth of biofilms in the labo-
ratory. The structure of the resulting biofilm,
especially the proportions of yeast-form and
hyphal-form cells, is strongly influenced by
parameters such as medium composition,
temperature, and the nature of the substratum
(59). Mutants unable to form hyphae or yeast-
form cells can nevertheless produce a biofilm,
demonstrating that a specific morphology is
not strictly essential for biofilm formation
(10). Because the proportions of the differ-
ent morphological forms vary depending on
environmental conditions, biofilm structure is
highly adaptable.
The content of exopolymeric matrix is also
influenced by biofilm incubation conditions.
Biofilms incubated statically contain relatively
low amounts of matrix, whereas biofilms in-
116 Kumamoto · Vinces
cubated with shaking produce higher lev-
els of matrix (42). This effect of medium
flow is also seen with bacterial biofilms (28).
Thus, flow of the medium above the sur-
face is another variable that influences biofilm
structure.
Quorum sensing also regulates biofilm for-
mation. The quorum-sensing molecule far-
nesol is produced continuously by growing
C. albicans cells, accumulating to levels corre-
lated with cell number, and acts as an inhibitor
of germination, i.e., the yeast-to-hypha tran-
sition (44). Incubation of cells in the presence
of farnesol leads to reduced biofilm formation
(86). Farnesol also has deleterious effects on
mature biofilms (86), suggesting that farnesol
regulates biofilm stability as well as biofilm
formation. Similar effects of quorum sensing
on mature bacterial biofilms have been noted
(106).
A two-component histidine kinase, Chk1p,
plays a role in the response of cells to far-
nesol. chk1 mutant cells are insensitive to the
effects of farnesol on both germination and
biofilm formation (58). Chk1p is a cytoplas-
mic protein that probably functions together
with currently unknown proteins to detect the
farnesol signal and produce the appropriate
response.
Thus, biofilms vary in the morphology and
organization of their cells and in their con-
tent of extracellular matrix. The plasticity in
biofilm structure in response to medium com-
position, the substratum, flow conditions, and
quorum sensing suggests that biofilms located
in different sites within the host or in associa-
tion with different types of devices or surfaces
are likely to differ in their properties.
Multiple Mechanisms Contribute
to the High Drug Resistance
of Biofilm Cells
One of the most vexing characteristics of
biofilms is their high-level resistance to an-
timicrobial drugs (28, 29, 59). Drug resistance
reflects a property of individual biofilm cells,
since C. albicans cells released from disrupted
 ANRV253-MI59-06 ARI 4 August 2005 11:16
biofilms still exhibit substantial levels of resis-
tance (8, 9, 85). In C. albicans biofilms, resis-
tance is probably not due to poor penetration
of drugs into the biofilm structure (1); mu-
tants that form structurally defective biofilms
nonetheless exhibit high drug resistance (87).
In the early stages of biofilm forma-
tion, changes in gene expression contribute
to resistance. Resistance to the antifungal
fluconazole can be detected as soon as 2 h
after adherence of C. albicans cells (71). At
this early time point, resistance is strongly de-
pendent on MDR1, which encodes a flucona-
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zole efflux facilitator. That is, an mdr1 null
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mutant biofilm is 16-fold more sensitive to
fluconazole than is a wild-type biofilm. Dele-
tion of CDR1 and CDR2, which encode ho-
mologous drug efflux pumps, also decreases
the resistance of adherent cells (71). Simi-
lar results were obtained after 6 h of ad-
herent growth, and increased efflux activity
in biofilm cells was detected (74). Thus, the
drug-resistant phenotype of biofilm cells at
early times after adherence results from ex-
pression of drug efflux determinants. Over-
expression of MDR1, CDR1, and CDR2 is
also important for drug resistance in drug-
resistant clinical strains (83).
In mature biofilms (e.g., after 48 h of incu-
bation) the mechanisms that confer drug re-
sistance are different. Mature biofilms formed
from mdr1 cdr1 cdr2 triple null mutants
or the various single and double mutants
are highly resistant to drugs (74, 85). Con-
sistent with these observations, expression
of the drug efflux determinants in mature
biofilms is not high (34, 74) and efflux ac-
tivity of mature biofilms is not higher than
the activity in planktonic cells (74). These
results argue against a role for the efflux
determinants in drug resistance in mature
biofilms.
Decreased membrane ergosterol content
(74) and altered expression of ergosterol
biosynthetic genes (34) in mature biofilm cells
have been noted. Therefore, increased drug
resistance in mature biofilms may be related
to an alteration in membrane sterol content.
Altered membrane composition could result
in changes in membrane properties such as
decreased permeability to drugs, leading to
higher drug resistance.
An alternative model has proposed that
most Pseudomonas aeruginosa cells in a
biofilm exhibit similar antibiotic resistance as
stationary-phase planktonic cells but that the
biofilm also contains a population of highly
resistant “persister” cells. The persister cells
are not mutants but rather wild-type cells
whose physiological state allows them to sur-
vive antibiotic treatment (64, 105). By sur-
viving antibiotic treatment, the persister cells
allow recovery of the population after an-
tibiotic treatment is discontinued. Although
the existence of persister cells in C. albicans
biofilms has not been tested directly, there is
evidence of cellular heterogeneity in C. al-
bicans biofilms (108), and thus this mecha-
nism could also contribute to biofilm drug
resistance.
In summary, multiple mechanisms con-
tribute to the increased drug resistance ex-
hibited by biofilms. Because drug resistance
mechanisms seen in early biofilms differ
from those conferring resistance in mature
biofilms, strategies designed to block early
biofilm formation may differ from strategies
that would be effective for eliminating mature
biofilms.
High Levels of Amino Acid
Biosynthesis and Protein Synthesis
in Biofilm Cells
To understand the distinctive biofilm lifestyle
at the molecular level, microarray studies of
gene expression in C. albicans biofilm cells
have been conducted (34). Results revealed
that, in comparison to postlogarithmic plank-
tonic cells that had been grown for the
same length of time, cells in biofilms express
higher levels of genes involved in amino acid
and nucleotide metabolism, protein synthe-
sis, other metabolic functions, and subcellular
localization. Garcia-Sanchez et al. (34) pro-
posed that biofilms require high-level protein
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biosynthesis, necessitating increased amino
acid biosynthesis.
Interestingly, analyses of gene expression
or protein composition in bacterial biofilm
cells compared with postlogarithmic plank-
tonic cells grown for the same length of time
showed some similarities to the results ob-
tained with C. albicans biofilms (98, 111).
Among the proteins or transcripts of genes
present at higher concentration in bacterial
biofilm cells were several gene products in-
volved in the synthesis of amino acids and
nucleotides or in protein translation. How-
ever, when bacterial biofilm cells were com-
pared with planktonic cells that were in the
exponential growth phase, these differences
in gene expression were not observed (98).
These results imply that biofilms contain cells
whose protein synthesis machinery is func-
tioning at the level seen in exponential-phase
cells, even though the biofilm has been devel-
oping for a long period.
Defective Biofilm Development
Caused by Mutations that
Alter the Cell Surface and
Compromise Adherence
Several mutations that reduce biofilm devel-
opment by compromising the first step in
biofilm formation, adherence to surfaces, have
been studied. One of these mutations affects
EFG1, a major regulator of hyphal develop-
ment (107). Efg1p regulates numerous genes
whose products include many cell surface pro-
teins (62, 77, 103). As a result of the cell
surface defect and the defect in production
of hyphae, it is not surprising that efg1 null
mutants are defective in biofilm development
(87). On polystyrene, the defect is evident
during the earliest stages of biofilm develop-
ment, i.e., adherence and microcolony forma-
tion (87). efg1 null mutants are also defective
in adhering to polyurethane central venous
catheter material (65). The EAP1 gene, which
encodes a predicted cell wall, GPI-anchored
protein, is dependent on Efg1p for expres-
sion and may be a critical target of Efg1p.
118 Kumamoto · Vinces
Expression of EAP1 in nonadherent Saccha-
romyces cerevisiae cells resulted in increased ad-
herence to polystyrene, suggesting that this
surface protein can mediate attachment to
polystyrene (66). Therefore, the failure of
efg1 mutants to initiate biofilm formation on
polystyrene may reflect the absence of Eap1p.
Other mutations affect adherence and
biofilm formation. Mutants lacking ACE2,
which encodes a transcription factor that
regulates expression of chitinase and cell
wall proteins, exhibit reduced adherence to
polystyrene and reduced biofilm formation
(50). Mutants lacking the NOT4 gene, which
encodes a putative E3 ubiquitin ligase, fail to
attach firmly to a serum-coated plastic surface
and are defective in biofilm formation (57). As
with the efg1 mutant, the absence of Not4p
or Ace2p may alter the surface properties of
C. albicans, leading to the observed defects in
adherence and biofilm development.
Adherence of Candida glabrata leading to
biofilm formation is mediated in part by
Epa6p, a cell surface protein. Expression of
EPA6 is activated under conditions that lead to
biofilm development through alteration of the
subtelomeric silencing machinery (48). Taken
together, these results demonstrate that alter-
ation of the cell surface by any one of several
mechanisms results in reduced adherence and
reduced biofilm development.
Development of Animal Models
for Biofilms
Although model systems make analysis of the
structure and development of biofilms acces-
sible to study, animal models are necessary
to understand the pathogenesis of biofilm-
related disease. Two animal models have been
recently described, one utilizing rabbits (100)
and one using rats (5). Both systems involve
placement of a central venous catheter fol-
lowed by direct inoculation of C. albicans cells
into the lumen of the catheter. The resulting
biofilms are structurally similar to biofilms de-
scribed in laboratory model systems, except
for the possible presence of host cells in the
 ANRV253-MI59-06 ARI 4 August 2005 11:16
biofilm (5). The catheter biofilms also exhibit
drug resistance and express the CDR2 gene,
consistent with results obtained in laboratory
model systems (5, 100). In the rat model,
biofilm development leads to seeding of the
kidneys with C. albicans, demonstrating dis-
seminated disease. These animal models offer
the potential for evaluation of new treatments
for biofilm infections based on insights from
laboratory model systems.
In summary, recent developments have
revealed molecular activities required for
biofilm development. Expression of appropri-
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ate cell surface molecules for adhesion and
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expression of genes needed for high levels
of protein synthesis are critical for successful
biofilm formation. Mechanisms underlying
antifungal resistance differ in early biofilms
and mature biofilms, indicating that different
strategies are needed to circumvent this prop-
erty of biofilms.
MECHANISMS OF INVASIVE
GROWTH
Tissue Invasion by C. albicans
Hyphae Occurs on Epithelial,
Epidermal, and Endothelial
Surfaces During Candidiasis
In superficial candidiasis, epidermal or epithe-
lial surface invasion by C. albicans hyphae is
commonly observed (79). When fungi colo-
nize an epithelial or epidermal surface, they
adhere to host cells and create depressions
in the surface of the host cells (45, 51, 60,
88). Fungal yeast-form cells also convert to
filamentous hyphae, which penetrate into the
surface.
During invasion and traversal of an
endothelial surface, the initial entry of yeast-
form cells into endothelial cells is by an endo-
cytotic process (22). Damage to the endothe-
lial surface (33, 52) results in exposure of the
underlying basement membrane, which may
then be invaded by hyphal-form cells.
In specimens scraped from human oral or
cutaneous lesions, most C. albicans cells are
found within host cells (20, 70, 73). Access to
the interior of host cells is probably achieved
by a combination of enzymatic activities (e.g.,
Endothelial:
proteases and phospholipase) and mechani- pertaining to tissue
cal force. Ultrastructural observations reveal composed of a layer
areas of clearing around penetrating hyphae, of flat squamous
supporting a role for lytic enzymes during in- cells, such as those
lining blood and
vasion (73, 99, 109). In addition, in samples
lymphatic vessels and
collected from cutaneous candidiasis cases, the heart
the corneocytes appear deformed as a result
of their interactions with C. albicans, support-
ing the notion that C. albicans uses mechani-
cal force to aid penetration (99). Studies with
model systems using explanted tissue, animal
infection, or cells in culture confirm these fea-
tures of invasion by C. albicans and demon-
strate that C. albicans hyphae may enter or pass
completely through host epithelial or epider-
mal cells with minimal damage to the host cell
(16, 32, 46, 88, 114).
In summary, hyphal penetration is a key
component of invasive growth, the process of
penetrating the substratum. Invasion of the
epithelial surface allows infecting organisms
to reach the bloodstream, and penetration and
disruption of endothelial surfaces allow or-
ganisms to escape from the bloodstream and
infect deep tissues. Thus, the invasive behav-
ior of C. albicans on a biological surface con-
tributes to disease pathogenesis.
Protease and Phospholipase
Activities Contribute to
Invasion of Host Tissue
Degradative enzymes secreted by C. albicans
cells are important for tissue invasion. Inhibi-
tion of the secreted aspartyl protease (SAP)
class of enzymes with the inhibitor pep-
statin decreases tissue invasion (76). Pepstatin
treatment also enhances survival following
intranasal challenge in mice by C. albicans
(31). In addition, several mutants lacking SAP
genes showed decreased invasion (76). Simi-
larly, secreted phospholipase activity has been
implicated in tissue invasion. In invading hy-
phae, phospholipase activity is concentrated
at the hyphal tip (35, 84); a mutant lacking
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 ANRV253-MI59-06 ARI 4 August 2005 11:16
secreted phospholipase B1 exhibits reduced
ability to penetrate cell monolayers (63) or
tissue (75). These data strongly implicate se-
creted proteases and phospholipases in suc-
cessful invasion.
Filamentation During Invasive
Growth is Promoted by Physical
Contact
As discussed above, hyphae that are capa-
ble of exerting mechanical force on host
cells are characteristically seen in specimens
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taken from infected tissue and probably con-
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tribute to invasion. Several experimental reg-
imens stimulate hyphal growth in the labora-
tory including the use of special media, high
temperature, or microaerophilic conditions
(30, 39).
To study invasive hyphal development
specifically, we have used an agar model sys-
tem that mimics some of the features of tissue
invasion. Growth of C. albicans cells embedded
Figure 1
Invasive growth of C. albicans within host tissue. Sections of fixed tongue
tissue from gnotobiotic immunosuppressed piglets orally inoculated with
(panel a) wild-type C. albicans or (panel b) efg1 cph1 double null mutant
cells. Immunohistochemical analysis with anti-C. albicans antibody is
shown. From Reference 91 with permission.
120 Kumamoto · Vinces
within agar medium stimulates the conversion
of yeast-form cells to filaments that invade the
medium (17). Control experiments indicate
that the important cue for hyphal production
is not reduced oxygen levels or gradients of
nutrients but rather physical contact of cells
with agar or other matrix material (17). Unlike
hyphal development stimulated by other cues,
invasive filamentation of cells in agar medium
is readily observed in rich medium at low or
high temperature.
Studies of immunosuppressed gnotobi-
otic piglets orally inoculated with C. albicans
suggest that contact-dependent filamentation
contributes to invasion of tissue. The efg1/efg1
cph1/cph1 double mutant strain, which lacks
two transcription factors that regulate fila-
mentation, forms filaments and invades the
tongue of the piglet (91) (Figure 1) or agar
medium (36, 91), but it is defective in fil-
amentation under all other conditions (68).
These results suggest that filamentation in re-
sponse to contact with a surface contributes
to invasion of epithelial tissue and is de-
pendent on factors other than Efg1p and
Cph1p.
Transcription Factors Regulating
Invasive Growth by Embedded Cells
To date, many genes that differentially affect
filamentous growth depending on whether
cells are grown in liquid or on agar medium
have been identified. These genes and their
phenotypes are summarized in Table 1. To
focus specifically on the molecular mecha-
nisms underlying the filamentous response
to growth within agar medium, the follow-
ing sections summarize the results of stud-
ies that make use of the embedded growth
condition (suspending cells within rich agar
medium).
Of the genes known to regulate inva-
sive growth, several are thought to encode
transcription factors (Table 1). The putative
transcription factor CZF1 plays a key role in
regulating the response of cells to embed-
ded conditions. Deletion of CZF1 results in
 ANRV253-MI59-06 ARI 4 August 2005 11:16

Table 1 C. albicans genes influencing filamentous growth within or on agar media

Gene Filamentation phenotype Putative function Reference(s)
Transcription factors
CPH1 Defective on Spider and SLAD Transcription factor 24, 67
agar; mild defect within agar
CZF1 Defective within agar Transcription factor 17
EFG1 Hyperfilamentous within agar Transcription factor 36
EFH1 efh1 efg1 double mutant Transcription factor 27
hyperfilamentous within agar
MCM1 Hyperfilamentous when Transcription factor 93
overexpressed on agar
Annu. Rev. Microbiol. 2005.59:113-33. Downloaded from arjournals.annualreviews.org

Signaling components
by Universidade Federal de Santa Catarina on 08/07/10. For personal use only.

CEK1 Defective on Lee’s and Spider agar MAPK 24, 53

CHK1 Defective on serum agar Histidine kinase 117
COS1 Defective on serum and Spider agar Histidine kinase 2, 117
CPP1 Hyperfilamentous and Phosphatase 23
hyperinvasive on agar
CST20 Defective on Spider agar MAPKKK kinase 24, 53
GAP1 Defective on solid Spider and Amino acid permease 12
SLAD
GPA2 Defective within agar G-protein α-subunit 69a, 72, 96
GPR1 Defective within agar G-protein-coupled receptor 69a, 72
HOG1 Defective in liquid and agar; MAPK 3
hyperinvasive on SLAD agar
HST7 Defective on Spider and SLAD MAPK kinase 24, 53, 96
agar; hyperfilamentous when
overexpressed within agar
RAS2 Active allele and mutant defective Small G-protein 69a, 96
within agar
SLN1 Defective on serum agar Histidine kinase 117
SSK1 Defective on agar; hyperinvasive on Two-component response regulator 19
SLAD agar
TPK1 Defective on agar PKA catalytic subunit 15
TPK2 Defect in invasive growth of yeast PKA catalytic subunit 15
cells on agar
Other genes
BMH1 Defective in liquid and within agar; 14-3-3 protein 92
one allele defective only in liquid
FAB1 Defective on Spider and serum agar Phosphatidylinositol 3-phosphate 7
5-kinase
PLD1 Defective on Spider agar; Phosphatidylcholine-specific 47
hyperfilamentous on phospholipase D1
cornmeal/Tween agar
RBR1 Defective on acidic M-199 soft agar pH-regulated putative cell wall 69
protein

www.annualreviews.org • C. albicans Growth on Surfaces 121

 ANRV253-MI59-06 ARI 4 August 2005 11:16

reduced invasive filamentation. Ectopic CZF1
expression results in accelerated filamenta-
tion (17) (Figure 2). Czf1p does not affect
MAPK:
mitogen-activated morphogenesis under other conditions, sug-
protein kinase gesting that a unique pathway is stimulated in
PKA: protein kinase embedded cells.
A Mutation of CPH1, the C. albicans homo-
logue of the S. cerevisiae STE12 transcription
factor, results in defective filamentous growth
in certain media (24, 62, 67) and mildly de-
fective filamentous growth within agar (36).
The czf1 cph1 double null mutant is more de-
fective than either single mutant, indicating
Annu. Rev. Microbiol. 2005.59:113-33. Downloaded from arjournals.annualreviews.org
mutation are epistatic to the effects of the czf1
mutation and partially epistatic to the effects
of the cph1 mutation (36), and therefore, the
efg1 cph1 double null mutant filaments during
growth within agar medium, as noted above.
CZF1 affects filamentation during growth
in agar (17). The effects of a CZF1 mutation,
however, are not observed in the absence of
EFG1 (36), which suggests that CZF1 pro-
motes filamentation by antagonizing EFG1-
mediated repression. Yeast two-hybrid data
suggest that this effect may involve phys-
ical interaction between Czf1p and Efg1p

that the two genes have partially overlapping (36).

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functions (17). The transcription factors Cph1p and

EFG1 encodes a basic helix-loop-helix
transcription factor that is required for hy-
phal development under most laboratory con-
ditions (68, 107). However, C. albicans efg1
mutant cells are more filamentous than wild-
type cells when embedded (36) (Figure 2a,b).
Thus, under embedded conditions, EFG1 acts
as a negative regulator of filamentous growth.
The related EFH1 gene is also believed to play
a negative role in regulating filamentation in
the embedded condition, perhaps in conjunc-
tion with Efg1p (27). The effects of the efg1
Efg1p each define two distinct signaling
pathways that regulate filamentous growth
under many conditions (30). Cph1p is regu-
lated by a MAPK cascade, and Efg1p is reg-
ulated by the cAMP/PKA signaling cascade.
The studies described above reveal the exis-
tence of at least two different programs in
which these cascades regulate morphogene-
sis (Figure 3). Under the standard condi-
tions used to promote filamentous growth,
e.g., the use of inducers such as serum, neutral
pH, microaerophilic conditions, temperature,

Figure 2
Invasive growth of C. albicans within agar medium. Colonies were grown on the surface of agar medium,
surface cells were washed off, and the remaining cells that were embedded within the agar were
photographed from the side. White arrowhead shows the top of the agar. Panel a, WT cells; panel b, efg1
null mutant cells; panel c, czf1 null mutant cells; and panel d, cells ectopically expressing CZF1 from the
promoter of the C. albicans maltase gene.

122 Kumamoto · Vinces

 ANRV253-MI59-06 ARI 4 August 2005 11:16
Annu. Rev. Microbiol. 2005.59:113-33. Downloaded from arjournals.annualreviews.org
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Figure 3
The standard and embedded programs for regulation of filamentation. Filamentation stimulated by
standard inducing conditions (e.g., serum, 37◦ C, neutral pH, starvation, microaerophilic growth) or by
embedded growth requires many of the same signaling components and transcription factors. Notable
differences include (a) the role of Efg1p as a positive regulator in the standard program and as a negative
regulator in the embedded program, and (b) a Czf1p-dependent pathway that promotes filamentation
only in the embedded program. Bold arrows indicate the relatively greater importance of the
Efg1-mediated pathways in both programs. Dashed lines indicate unclear relationships. Blunt arrows
indicate negative effects. Transcription factors are depicted as boxes. For simplicity, many factors with
uncertain relationships to these pathways have been left out of the figure but are discussed in greater
detail in the text.

and nutritional signals (30, 39), hyphal de- Signaling Pathways Regulating
velopment by the standard program requires Invasive Growth by Embedded Cells
Efg1p as a positive regulator. Cph1p performs
The C. albicans genes GPR1, GPA2, and RAS2
a backup, positive function in the standard
regulate filamentous growth under embed-
program. During embedded growth, filamen-
ded conditions (69a, 72, 96). GPR1, which
tation is promoted by contact with agar or
encodes a G-protein-coupled receptor, and
other matrix material. Filamentation under
GPA2, which encodes a G-protein α-subunit
embedded conditions is controlled by the em-
homologue, probably function in transmis-
bedded program, in which Efg1p acts as a
sion of signals. gpr1 and gpa2 mutants are
negative regulator and Czf1p functions as an
defective in filamentous growth under vari-
antagonist of Efg1p repressor activity. The
ous conditions, particularly during embedded
function of Cph1p partially overlaps that of
growth (69a, 72, 96). Recent studies suggest
Czf1p in the embedded program. The po-
that GPR1 and GPA2 act in the cAMP-PKA
tential contributions of other transcriptional
pathway (69a, 72). These studies demonstrate
regulators of morphogenesis (25) to regula-
suppression of the gpa2 defect by addition
tion in the embedded condition are currently
of exogenous cAMP. In addition, the gpa2
unknown.

www.annualreviews.org • C. albicans Growth on Surfaces 123

 ANRV253-MI59-06 ARI 4 August 2005 11:16
mutation is rescued by overexpression of
TPK1, a component of the cAMP-PKA path-
way, but not by overexpression of HST7, a
member of the MAPK pathway (69a).
Under standard inducing conditions, the
small G-protein-encoding RAS1 gene posi-
tively regulates filamentous growth and is be-
lieved to act in both the cAMP and MAPK
pathways (62a) (Figure 3). Deletion of RAS1
causes a defect in filamentation under em-
bedded conditions, and this defect can be
suppressed by addition of exogenous cAMP,
suggesting a positive role for both RAS1 and
Annu. Rev. Microbiol. 2005.59:113-33. Downloaded from arjournals.annualreviews.org
cAMP in regulating filamentous growth in
by Universidade Federal de Santa Catarina on 08/07/10. For personal use only.
this condition (69a). Although under stan-
dard conditions exogenous cAMP acceler-
ates filamentous growth, the influence of
cAMP on filamentation of embedded cells is
unclear.
Taken together, the data suggest that under
embedded conditions the MAPK pathway ac-
tivates a positive regulator, Cph1p, (53) and
promotes filamentous growth, whereas the
cAMP-dependent pathway, including RAS1,
GPR1, and GPA2, acts on a negative regula-
tor, Efg1p, (14) that suppresses filamentous
growth. The signaling components that lie
upstream of Czf1p are not yet known.
The two genes encoding isoforms of cat-
alytic subunits of PKA, a component of the
cAMP signaling pathway, have distinct roles
in filamentation in liquid and agar media (15).
The tpk1 mutant is defective for filamentous
growth on agar media and is only slightly
affected in liquid, and the tpk2 mutant is
only partially defective on agar but is strongly
blocked for filamentation in liquid. The basis
for the differential effects of TPK1 and TPK2
is currently unclear.
Other genes that affect morphogenesis
on agar medium include SLN1, COS1, and
CHK1, which encode two-component histi-
dine kinases, SSK1, which encodes a response
regulator, and HOG1, which encodes an os-
moregulating MAPK gene (2, 3, 19, 117).
These genes may not act within the MAPK
or cAMP pathways but rather within indepen-
dent pathways.
124 Kumamoto · Vinces
The activities of numerous other genes
that specifically affect filamentation on agar
medium have been described, but the mech-
anisms by which they act are incompletely
understood. These genes are summarized in
Table 1.
Models for Invasion of Tissue
Surfaces
Studies of the efg1 and/or the efg1 cph1 dou-
ble null mutant in different animal models of
infection reveal two mechanisms for penetra-
tion of tissue surfaces. One mechanism occurs
on mucosal surfaces and involves Efg1p-
dependent adherence and proliferation on the
surface, followed by Efg1p-independent pen-
etration of the surface. A second mechanism
occurs on endothelial surfaces and involves
uptake of fungi by endocytosis, followed by
intracellular, Efg1p-dependent hyphal devel-
opment that allows fungal penetration of the
surface.
The extensive surface growth of wild-type
C. albicans in pseudomembranous oral can-
didiasis in the immunosuppressed gnotobiotic
piglet is dependent on Efg1p and is not ob-
served with the efg1 cph1 double null mutant
(6). In addition, defects in adhesion of efg1 null
mutant or efg1 cph1 double null mutant cells
to oral epithelial cells and Caco-2 cells have
been observed (26, 112).
In several systems, the overall extent of in-
vasion is reduced in the absence of Efg1p,
probably reflecting defective adherence (26,
40, 56, 114). However, invasion of host ep-
ithelium by C. albicans can occur in the ab-
sence of Efg1p (91) (Figure 1b). In fact,
invasive growth probably requires downregu-
lation of EFG1 function. This model is based
on the observations that efg1 null mutants are
hyperfilamentous and hyperinvasive in agar
(36) (Figure 2a,b) and that EFG1 transcrip-
tion is downregulated during filamentation in
liquid medium (107, 110).
Therefore, because Czf1p is a negative
regulator of Efg1p (36), Czf1p may act as a
switch that inactivates Efg1p and promotes
 ANRV253-MI59-06 ARI 4 August 2005 11:16
the conversion from surface growth to inva-
sive growth. Activation of the Czf1p switch
occurs as a response to contact with agar or
other matrix material. Czf1p antagonism of
Efg1p is not the only mechanism that pro-
motes invasive filamentation, since there are
also high-temperature-activated mechanisms
that do not require Czf1p (17). The result of
Efg1p and Czf1p activities is the characteris-
tic invasion of epithelial surfaces seen in pseu-
domembranous oral candidiasis.
In contrast, other host cell types such
as endothelial cells are stimulated to take
Annu. Rev. Microbiol. 2005.59:113-33. Downloaded from arjournals.annualreviews.org
up C. albicans cells by endocytosis, and thus
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a second mechanism for invasion is promi-
nent on these surfaces. In this situation,
invasion is initiated by endocytosis of yeast-
form cells. Within host cells, the fungi un-
dergo Efg1p-dependent formation of germ
tubes and exit from the host cell. Germina-
tion within host cells is defective when efg1
mutant cells are internalized by macrophages
(68) or neutrophils (56). Defective exit by ger-
mination and defective endocytosis probably
contribute to the failure of efg1 mutants to in-
vade and damage endothelial cell monolayers
(81).
In summary, tissue invasion requires
degradative activities secreted by the invad-
ing C. albicans hyphae and mechanical forces
exerted by the hyphae. The mechanisms
that regulate invasion are adapted to the
nature of the surface to which C. albicans cells
are bound. The mechanism for invasion of
mucosal surfaces resembles the embedded
program for regulation of filamentation. That
is, following Efg1p-dependent proliferation
on the surface, Efg1p is downregulated and
contact-dependent hyphal development en-
sues. A second mechanism involving entry of
fungi into host cells via endocytosis followed
by intracellular, Efg1p-dependent filamenta-
tion and exit from the host cells is important
on endothelial surfaces. In this situation,
control of hyphal development resembles the
standard program for regulation of filamen-
tation. The cues that promote filamentation
on the two types of surfaces are different and
the molecules involved in controlling hyphal
morphogenesis are used in different ways.
The existence of the two mechanisms for
filamentation contributes to the remarkable
versatility of C. albicans as a pathogen.
GUIDANCE OF HYPHAE BY
THIGMOTROPISM
The direction of C. albicans hyphal growth
can be determined by the topography of
the substratum. This behavior, known as
thigmotropism, allows hyphae to be guided
by ridges in the substratum (113). In stud-
ies of this behavior, hyphae penetrate the
pores of nucleopore membranes and, after
penetration, follow the face of the mem-
brane. During penetration, hyphae grow
both away from and toward the agar un-
derneath the membrane, implying that the
orientation of hyphal growth is not due
to chemotropism but to thigmotropism (38,
101). Helical growth of hyphae occurs when
cells are grown on firm surfaces, such as cel-
lophane (102). These thigmotropic responses
to surface features are reminiscent of the
behavior of plant-pathogenic fungi on leaf
surfaces.
Thigmotropism plays a major role in the
location of infectable sites on plants by phy-
topathogenic fungi. For example, Uromyces
appendiculatus, the causative agent of bean
rust, produces hyphae that are guided by
the topography of the bean leaf surface to
grow toward stomata, where they differen-
tiate into appressoria, specialized structures
needed for invasion of the leaf. Induction of
appressorium formation is caused by archi-
tectural features of the stomatal guard cells
and can be mimicked in the laboratory us-
ing polystyrene membranes bearing ridges of
the appropriate height. The plant pathogens
Puccinia hordei and Magnaporthe grisea also use
thigmotropic behavior on the surface of plants
to locate openings in the epidermal layer of
plant tissue and initiate invasive growth (43,
89, 116). Hence, thigmotropism plays an im-
portant role in guidance of hyphal growth,
www.annualreviews.org • C. albicans Growth on Surfaces 125
 ANRV253-MI59-06 ARI 4 August 2005 11:16

in regulation of development, and in disease To summarize, thigmotropic behavior al-

progression.
Thigmotropism has also been demon-
MS:
mechanosensitive strated in dermatophytes and in saprophytes
such as Mucor mucedo and Neurospora crassa
(80). Thus, thigmotropic behavior of hyphae
is not restricted to pathogenic fungi; it is a
general feature of fungal hyphae that must
forage for nutrients on surfaces and within
materials.
The thigmotropic response of C. albicans
may be an adaptation for penetrating tissue.
Studies of cultured intestinal enterocytes
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lows C. albicans to respond with great
precision to topographical features of the
surface. This behavior may contribute to
tissue invasion by directing penetrating
hyphae toward more vulnerable regions of
the tissue. Further studies will illuminate the
details of the molecular mechanisms used
by C. albicans to sense the features of its
environment.
CONCLUSIONS

infected with C. albicans (115) and of biop- Contact with surfaces elicits unique physio-
by Universidade Federal de Santa Catarina on 08/07/10. For personal use only.

sies of oral candidiasis taken from AIDS
patients (90) demonstrated orientation of hy-
phae along ridges or grooves and penetration
into the regions between enterocytes or cor-
neocytes. The authors suggested that thig-
motropism may allow hyphae to locate the
intercellular regions.
MS ion channels are found in organisms
from archeans, E. coli, and yeast to higher eu-
karyotes (13, 49, 55, 118) and are believed
to play roles in processes such as osmosensa-
tion, topographic sensing, touch, and hearing.
MS channels are activated by stretch forces
on the plasma membrane, resulting in the
opening of the channels. C. albicans possesses
MS channel activity. Reorientation of hyphal
growth in response to ridges is attenuated by
treatment with an inhibitor of MS channels,
Gd3+ , which suggests that MS channels are
responsible for sensing substrate topography
(113).
logical responses in C. albicans. On solid sur-
faces, C. albicans cells form biofilms with char-
acteristic three-dimensional structures and
high antifungal resistance. This response to
contact with a surface is significant for hu-
man health because of the large numbers
of implanted medical devices used in mod-
ern medicine. On tissue surfaces, C. al-
bicans initially adheres and grows on the
surface and subsequently produces invasive
filaments that penetrate the surface. This
behavior allows organisms to escape from
their normal niche in the gastrointestinal
tract and reach the bloodstream, setting
the stage for disseminated, life-threatening
infection. Understanding the unique physiol-
ogy of C. albicans on surfaces and the adap-
tations of the organism that favor infection,
researchers will be better equipped to develop
methods for interrupting the progression to
disease.

SUMMARY POINTS
1. Growth on surfaces is a natural part of the C. albicans lifestyle.
2. The unique physiology of cells growing on a surface makes an important contribution
to pathogenesis.
3. Biofilm formation, a behavioral response of C. albicans cells growing on a surface, is
associated with medical device infection.
4. The morphology and organization of C. albicans biofilms are influenced by numerous
parameters, and thus variability may be observed in biofilms located at different sites
within the host.

126 Kumamoto · Vinces

 ANRV253-MI59-06 ARI 4 August 2005 11:16

5. Multiple mechanisms contribute to the high resistance of biofilms to antifungal drugs.

6. Secreted degradative enzymes and the hyphal form of growth play key roles during
tissue invasion by C. albicans.
7. Mechanisms that promote invasion of epithelial surfaces differ from mechanisms used
for invasion of endothelial surfaces.
8. EFG1 regulates several aspects of growth on surfaces, including adhesion, biofilm
development, colonization, and invasion.

ACKNOWLEDGMENTS
Annu. Rev. Microbiol. 2005.59:113-33. Downloaded from arjournals.annualreviews.org

We are grateful to Perry Riggle, Julia Köhler, and Linc Sonenshein for careful reading of the
by Universidade Federal de Santa Catarina on 08/07/10. For personal use only.

manuscript and for helpful comments. Our research on this project was supported by grant
AI38591 from the National Institute of Allergy and Infectious Diseases.

LITERATURE CITED
1. Al-Fattani MA, Douglas LJ. 2004. Penetration of Candida biofilms by antifungal agents.
Antimicrob. Agents Chemother. 48:3291–97
2. Alex LA, Korch C, Selitrennikoff CP, Simon MI. 1998. COS1, a two-component histidine
kinase that is involved in hyphal development in the opportunistic pathogen Candida
albicans. Proc. Natl. Acad. Sci. USA 95:7069–73
3. Alonso-Monge R, Navarro-Garcia F, Molero G, Diez-Orejas R, Gustin M, et al. 1999.
Role of the mitogen-activated protein kinase Hog1p in morphogenesis and virulence of
Candida albicans. J. Bacteriol. 181:3058–68
4. Ampel NM. 1996. Emerging disease issues and fungal pathogens associated with HIV
infection. Emerg. Infect. Dis. 2:109–16
5. Andes D, Nett J, Oschel P, Albrecht R, Marchillo K, Pitula A. 2004. Development and
characterization of an in vivo central venous catheter Candida albicans biofilm model.
Infect. Immun. 72:6023–31
6. Andrutis KA, Riggle PJ, Kumamoto CA, Tzipori S. 2000. Intestinal lesions associated with
disseminated candidiasis in an experimental animal model. J. Clin. Microbiol. 38:2317–23
7. Augsten M, Hubner C, Nguyen M, Kunkel W, Hartl A, Eck R. 2002. Defective hyphal
induction of a Candida albicans phosphatidylinositol 3-phosphate 5-kinase null mutant on
solid media does not lead to decreased virulence. Infect. Immun. 70:4462–70
8. Baillie GS, Douglas LJ. 1998. Effect of growth rate on resistance of Candida albicans
This careful study
biofilms to antifungal agents. Antimicrob. Agents Chemother. 42:1900–5 of the resistance of
9. Baillie GS, Douglas LJ. 1998. Iron-limited biofilms of Candida albicans and their suscep- biofilm cells
tibility to amphotericin B. Antimicrob. Agents Chemother. 42:2146–49 demonstrates that
10. Baillie GS, Douglas LJ. 1999. Role of dimorphism in the development of Candida albicans resuspended cells
from a biofilm
biofilms. J. Med. Microbiol. 48:671–79
exhibit enhanced
11. Baillie GS, Douglas LJ. 2000. Matrix polymers of Candida biofilms and their possible role antifungal
in biofilm resistance to antifungal agents. J. Antimicrob. Chemother. 46:397–403 resistance.
12. Biswas S, Roy M, Datta A. 2003. N-acetylglucosamine-inducible CaGAP1 encodes a
general amino acid permease which co-ordinates external nitrogen source response and
morphogenesis in Candida albicans. Microbiology 149:2597–608

www.annualreviews.org • C. albicans Growth on Surfaces 127

 ANRV253-MI59-06 ARI 4 August 2005 11:16

13. Blount P. 2003. Molecular mechanisms of mechanosensation: big lessons from small cells.
Neuron 37:731–34
14. Bockmühl DP, Ernst JF. 2001. A potential phosphorylation site for an A-type kinase
in the Efg1 regulator protein contributes to hyphal morphogenesis of Candida albicans.
Genetics 157:1523–30
15. Bockmühl DP, Krishnamurthy S, Gerads M, Sonneborn A, Ernst JF. 2001. Distinct and
redundant roles of the two protein kinase A isoforms Tpk1p and Tpk2p in morphogenesis
and growth of Candida albicans. Mol. Microbiol. 42:1243–57
16. Borg M, Ruchel R. 1988. Expression of extracellular acid proteinase by proteolytic Candida
spp. during experimental infection of oral mucosa. Infect. Immun. 56:626–31
17. Brown DH Jr, Giusani AD, Chen X, Kumamoto CA. 1999. Filamentous growth of
The authors
demonstrate that Candida albicans in response to physical environmental cues, and its regulation by
growth within agar the unique CZF1 gene. Mol. Microbiol. 34:651–62
Annu. Rev. Microbiol. 2005.59:113-33. Downloaded from arjournals.annualreviews.org

promotes 18. Calderone RA. 2002. Introduction and historical perspectives. In Candida and Candidiasis,
by Universidade Federal de Santa Catarina on 08/07/10. For personal use only.

filamentation and ed. RA Calderone, pp. 3–13. Washington, DC: ASM Press
that CZF1 is an
19. Calera JA, Zhao XJ, Calderone R. 2000. Defective hyphal development and avirulence
important
regulator of this caused by a deletion of the SSK1 response regulator gene in Candida albicans. Infect. Immun.
response. 68:518–25
20. Cawson RA, Rajasingham KC. 1972. Ultrastructural features of the invasive phase of
Candida albicans. Br. J. Dermatol. 87:435–43
21. Chandra J, Kuhn DM, Mukherjee PK, Hoyer LL, McCormick T, Ghannoum MA.
This article
includes a study 2001. Biofilm formation by the fungal pathogen Candida albicans: development,
of the stages architecture, and drug resistance. J. Bacteriol. 183:5385–94
of biofilm 22. Clemons KV, Calich VL, Burger E, Filler SG, Grazziutti M, et al. 2000. Pathogenesis I:
development and interactions of host cells and fungi. Med. Mycol. 38(Suppl. 1):99–111
the architecture of
23. Csank C, Makris C, Meloche S, Schröppel K, Rollinghoff M, et al. 1997. Derepressed
the mature biofilm.
hyphal growth and reduced virulence in a VH1 family-related protein phosphatase mutant
of the human pathogen Candida albicans. Mol. Biol. Cell 8:2539–51
24. Csank C, Schröppel K, Leberer E, Harcus D, Mohamed O, et al. 1998. Roles of the Can-
dida albicans mitogen-activated protein kinase homolog, Cek1p, in hyphal development
and systemic candidiasis. Infect. Immun. 66:2713–21
25. Davis D. 2003. Adaptation to environmental pH in Candida albicans and its relation to
pathogenesis. Curr. Genet. 44:1–7
26. Dieterich C, Schandar M, Noll M, Johannes FJ, Brunner H, et al. 2002. In vitro re-
constructed human epithelia reveal contributions of Candida albicans EFG1 and CPH1 to
adhesion and invasion. Microbiology 148:497–506
27. Doedt T, Krishnamurthy S, Bockmühl DP, Tebarth B, Stempel C, et al. 2004. APSES pro-
teins regulate morphogenesis and metabolism in Candida albicans. Mol. Biol. Cell 15:3167–
80
28. Donlan RM, Costerton JW. 2002. Biofilms: survival mechanisms of clinically relevant
microorganisms. Clin. Microbiol. Rev. 15:167–93
29. Douglas LJ. 2003. Candida biofilms and their role in infection. Trends Microbiol. 11:30–36
30. Ernst JF. 2000. Transcription factors in Candida albicans: environmental control of mor-
phogenesis. Microbiology 146(Pt. 8):1763–74
31. Fallon K, Bausch K, Noonan J, Huguenel E, Tamburini P. 1997. Role of aspartic proteases
in disseminated Candida albicans infection in mice. Infect. Immun. 65:551–56
32. Farrell SM, Hawkins DF, Ryder TA. 1983. Scanning electron microscope study of Candida
albicans invasion of cultured human cervical epithelial cells. Sabouraudia 21:251–54

128 Kumamoto · Vinces

 ANRV253-MI59-06 ARI 4 August 2005 11:16

33. Filler SG, Swerdloff JN, Hobbs C, Luckett PM. 1995. Penetration and damage of en-
dothelial cells by Candida albicans. Infect. Immun. 63:976–83
34. Garcia-Sanchez S, Aubert S, Iraqui I, Janbon G, Ghigo JM, d’Enfert C. 2004. Candida
albicans biofilms: a developmental state associated with specific and stable gene expression
patterns. Eukaryot. Cell 3:536–45
35. Ghannoum MA. 2000. Potential role of phospholipases in virulence and fungal patho-
genesis. Clin. Microbiol. Rev. 13:122–43
36. Giusani AD, Vinces M, Kumamoto CA. 2002. Invasive filamentous growth of Candida
albicans is promoted by Czf1p-dependent relief of Efg1p-mediated repression. Genetics
160:1749–53
37. Gorbach SL, Nahas L, Lerner PI, Weinstein L. 1967. Studies of intestinal microflora. I.
Effects of diet, age, and periodic sampling on numbers of fecal microorganisms in man.
Gastroenterology 53:845–55
Annu. Rev. Microbiol. 2005.59:113-33. Downloaded from arjournals.annualreviews.org

38. Gow NA. 1994. Growth and guidance of the fungal hypha. Microbiology 140:3193–205
by Universidade Federal de Santa Catarina on 08/07/10. For personal use only.

39. Gow NA. 1997. Germ tube growth of Candida albicans. Curr. Top. Med. Mycol. 8:43–55
40. Gow NA, Knox Y, Munro CA, Thompson WD. 2003. Infection of chick chorioallantoic
membrane (CAM) as a model for invasive hyphal growth and pathogenesis of Candida
albicans. Med. Mycol. 41:331–38
41. Gudlaugsson O, Gillespie S, Lee K, Vande Berg J, Hu J, et al. 2003. Attributable mortality
of nosocomial candidemia, revisited. Clin. Infect. Dis. 37:1172–77
42. Hawser SP, Baillie GS, Douglas LJ. 1998. Production of extracellular matrix by Candida
albicans biofilms. J. Med. Microbiol. 47:253–56
43. Hoch HC, Staples RC, Whitehead B, Comeau J, Wolf ED. 1987. Signaling for growth
orientation and cell differentiation by surface topography in Uromyces. Science 235:1659–
62
44. Hornby JM, Jensen EC, Lisec AD, Tasto JJ, Jahnke B, et al. 2001. Quorum sensing in
the dimorphic fungus Candida albicans is mediated by farnesol. Appl. Environ. Microbiol.
67:2982–92
45. Hoshika K, Mine H. 1994. Significance of modes of adherence in esophageal Candida
albicans. J. Gastroenterol. 29:1–5
46. Howlett JA, Squier CA. 1980. Candida albicans ultrastructure: colonization and invasion
of oral epithelium. Infect. Immun. 29:252–60
47. Hube B, Hess D, Baker CA, Schaller M, Schafer W, Dolan JW. 2001. The role and
relevance of phospholipase D1 during growth and dimorphism of Candida albicans. Mi-
crobiology 147:879–89
48. Iraqui I, Garcia-Sanchez S, Aubert S, Dromer F, Ghigo JM, et al. 2005. The Yak1p
kinase controls expression of adhesins and biofilm formation in Candida glabrata in a
Sir4p dependent pathway. Mol. Microbiol. 55:1259–71
49. Kanzaki M, Nagasawa M, Kojima I, Sato C, Naruse K, et al. 1999. Molecular identifi-
cation of a eukaryotic, stretch-activated nonselective cation channel. Science 285:882–86.
Erratum. 1999. Science 285(5433):1493
50. Kelly MT, MacCallum DM, Clancy SD, Odds FC, Brown AJ, Butler G. 2004. The Can-
dida albicans CaACE2 gene affects morphogenesis, adherence and virulence. Mol. Microbiol.
53:969–83
51. Kennedy MJ, Volz PA, Edwards CA, Yancey RJ. 1987. Mechanisms of association of
Candida albicans with intestinal mucosa. J. Med. Microbiol. 24:333–41
52. Klotz SA, Drutz DJ, Harrison JL, Huppert M. 1983. Adherence and penetration of
vascular endothelium by Candida yeasts. Infect. Immun. 42:374–84

www.annualreviews.org • C. albicans Growth on Surfaces 129

 ANRV253-MI59-06 ARI 4 August 2005 11:16

53. Köhler JR, Fink GR. 1996. Candida albicans strains heterozygous and homozygous for mu-
tations in mitogen-activated protein kinase signaling components have defects in hyphal
development. Proc. Natl. Acad. Sci. USA 93:13223–28
54. Kojic EM, Darouiche RO. 2004. Candida infections of medical devices. Clin. Microbiol.
Rev. 17:255–67
55. Koprowski P, Kubalski A. 2001. Bacterial ion channels and their eukaryotic homologues.
Bioessays 23:1148–58
56. Korting HC, Hube B, Oberbauer S, Januschke E, Hamm G, et al. 2003. Reduced expres-
sion of the hyphal-independent Candida albicans proteinase genes SAP1 and SAP3 in the
efg1 mutant is associated with attenuated virulence during infection of oral epithelium. J.
Med. Microbiol. 52:623–32
57. Krueger KE, Ghosh AK, Krom BP, Cihlar RL. 2004. Deletion of the NOT4 gene impairs
hyphal development and pathogenicity in Candida albicans. Microbiology 150:229–40
Annu. Rev. Microbiol. 2005.59:113-33. Downloaded from arjournals.annualreviews.org

58. Kruppa M, Krom BP, Chauhan N, Bambach AV, Cihlar RL, Calderone RA. 2004. The
by Universidade Federal de Santa Catarina on 08/07/10. For personal use only.

two-component signal transduction protein Chk1p regulates quorum sensing in Candida

albicans. Eukaryot. Cell 3:1062–65
59. Kumamoto CA. 2002. Candida biofilms. Curr. Opin. Microbiol. 5:608–11
60. Kvaal C, Lachke SA, Srikantha T, Daniels K, McCoy J, Soll DR. 1999. Misexpression of
the opaque-phase-specific gene PEP1 (SAP1) in the white phase of Candida albicans confers
increased virulence in a mouse model of cutaneous infection. Infect. Immun. 67:6652–62
61. Lamfon H, Porter SR, McCullough M, Pratten J. 2003. Formation of Candida albicans
biofilms on non-shedding oral surfaces. Eur. J. Oral Sci. 111:465–71
62. Lane S, Birse C, Zhou S, Matson R, Liu H. 2001. DNA array studies demonstrate con-
vergent regulation of virulence factors by Cph1, Cph2, and Efg1 in Candida albicans. J.
Biol. Chem. 276:48988–96
62a. Leberer E, Harcus D, Dignard D, Johnson L, Ushinsky S, et al. 2001. Ras links cellu-
lar morphogenesis to virulence by regulation of the MAP kinase and cAMP signalling
pathways in the pathogenic fungus Candida albicans. Mol. Microbiol. 42:673–87
63. Leidich SD, Ibrahim AS, Fu Y, Koul A, Jessup C, et al. 1998. Cloning and disruption of
caPLB1, a phospholipase B gene involved in the pathogenicity of Candida albicans. J. Biol.
Chem. 273:26078–86
64. Lewis K. 2001. Riddle of biofilm resistance. Antimicrob. Agents Chemother. 45:999–1007
65. Lewis RE, Lo HJ, Raad II, Kontoyiannis DP. 2002. Lack of catheter infection by the
efg1/efg1 cph1/cph1 double-null mutant, a Candida albicans strain that is defective in fila-
mentous growth. Antimicrob. Agents Chemother. 46:1153–55
66. Li F, Palecek SP. 2003. EAP1, a Candida albicans gene involved in binding human epithelial
cells. Eukaryot. Cell 2:1266–73
67. Liu H, Köhler J, Fink GR. 1994. Suppression of hyphal formation in Candida albicans by
mutation of a STE12 homolog. Science 266:1723–26. Erratum. 1995. Science 267(5194):17
68. Lo H-J, Köhler JR, DiDomenico B, Loebenberg D, Cacciapuoti A, Fink GR. 1997.
This important
study demonstrates Nonfilamentous C. albicans mutants are avirulent. Cell 90:939–49
that a mutant 69. Lotz H, Sohn K, Brunner H, Mühlschlegel FA, Rupp S. 2004. RBR1, a novel pH-regulated
lacking both Efg1p cell wall gene of Candida albicans, is repressed by RIM101 and activated by NRG1. Eukaryot.
and Cph1p is Cell 3:776–84
nonfilamentous 69a. Maidan MM, De Rop L, Serneels J, Exler S, Rupp S, et al. 2005. The G protein-coupled
under almost all
receptor Gpr1 and the Gα protein Gpa2 act through the cAMP-PKA pathway to induce
conditions and is
avirulent. morphogenesis in Candida albicans. Mol. Biol. Cell (Epub ahead of print)
70. Marrie TJ, Costerton JW. 1981. The ultrastructure of Candida albicans infections. Can.
J. Microbiol. 27:1156–64

130 Kumamoto · Vinces

 ANRV253-MI59-06 ARI 4 August 2005 11:16

71. Mateus C, Crow SA Jr, Ahearn DG. 2004. Adherence of Candida albicans to silicone
induces immediate enhanced tolerance to fluconazole. Antimicrob. Agents Chemother. 48:
3358–66
72. Miwa T, Tagaki Y, Shinozaki M, Yun C-W, Schell WA, et al. 2004. Gpr1, a puta-
tive G-protein-coupled receptor, regulates morphogenesis and hypha formation in the
pathogenic fungus Candida albicans. Eukaryot. Cell 3:919–31
73. Montes LF, Wilborn WH. 1968. Ultrastructural features of host-parasite relationship in
oral candidiasis. J. Bacteriol. 96:1349–56
74. Mukherjee PK, Chandra J, Kuhn DM, Ghannoum MA. 2003. Mechanism of fluconazole
resistance in Candida albicans biofilms: phase-specific role of efflux pumps and membrane
sterols. Infect. Immun. 71:4333–40
75. Mukherjee PK, Seshan KR, Leidich SD, Chandra J, Cole GT, Ghannoum MA. 2001.
Reintroduction of the PLB1 gene into Candida albicans restores virulence in vivo. Micro-
Annu. Rev. Microbiol. 2005.59:113-33. Downloaded from arjournals.annualreviews.org

biology 147:2585–97
by Universidade Federal de Santa Catarina on 08/07/10. For personal use only.

76. Naglik JR, Challacombe SJ, Hube B. 2003. Candida albicans secreted aspartyl proteinases
in virulence and pathogenesis. Microbiol. Mol. Biol. Rev. 67:400–28
77. Nantel A, Dignard D, Bachewich C, Harcus D, Marcil A, et al. 2002. Transcription
profiling of Candida albicans cells undergoing the yeast-to-hyphal transition. Mol. Biol.
Cell 13:3452–65
78. Odds FC. 1987. Candida infections: an overview. Crit. Rev. Microbiol. 15:1–5
79. Odds FC. 1994. Pathogenesis of Candida infections. J. Am. Acad. Dermatol. 31:S2–5
80. Perera TH, Gregory DW, Marshall D, Gow NA. 1997. Contact-sensing by hyphae of
dermatophytic and saprophytic fungi. J. Med. Vet. Mycol. 35:289–93
81. Phan QT, Belanger PH, Filler SG. 2000. Role of hyphal formation in interactions of
Candida albicans with endothelial cells. Infect. Immun. 68:3485–90
82. Pope LM, Cole GT. 1981. SEM studies of adherence of Candida albicans to the gastroin-
testinal tract of infant mice. Scan Electron Microsc. Pt. 3:73–80
83. Prasad R, Panwar SL, Smriti. 2002. Drug resistance in yeasts: an emerging scenario. Adv.
Microb. Physiol. 46:155–201
84. Pugh D, Cawson RA. 1977. The cytochemical localization of phospholipase in Candida
albicans infecting the chick chorio-allantoic membrane. Sabouraudia 15:29–35
85. Ramage G, Bachmann S, Patterson TF, Wickes BL, Lopez-Ribot JL. 2002. Investigation
of multidrug efflux pumps in relation to fluconazole resistance in Candida albicans biofilms.
J. Antimicrob. Chemother. 49:973–80
86. Ramage G, Saville SP, Wickes BL, Lopez-Ribot JL. 2002. Inhibition of Candida albi-
cans biofilm formation by farnesol, a quorum-sensing molecule. Appl. Environ. Microbiol.
68:5459–63
87. Ramage G, VandeWalle K, Lopez-Ribot JL, Wickes BL. 2002. The filamentation pathway
controlled by the Efg1 regulator protein is required for normal biofilm formation and
development in Candida albicans. FEMS Microbiol. Lett. 214:95–100
88. Ray TL, Payne CD. 1988. Scanning electron microscopy of epidermal adherence and
cavitation in murine candidiasis: a role for Candida acid proteinase. Infect. Immun. 56:1942–
49
89. Read ND, Kellock LJ, Collins TJ, Gundlach AM. 1997. Role of topography sensing for
infection-structure differentiation in cereal rust fungi. Planta 202:163–70
90. Reichart PA, Philipsen HP, Schmidt-Westhausen A, Samaranayake LP. 1995. Pseu-
domembranous oral candidiasis in HIV infection: ultrastructural findings. J. Oral Pathol.
Med. 24:276–81

www.annualreviews.org • C. albicans Growth on Surfaces 131

 ANRV253-MI59-06 ARI 4 August 2005 11:16

91. Riggle PJ, Andrutis KA, Chen X, Tzipori SR, Kumamoto CA. 1999. Invasive lesions
containing filamentous forms produced by a Candida albicans mutant that is defective in
filamentous growth in culture. Infect. Immun. 67:3649–52
92. Roberts RL, Mosch HU, Fink GR. 1997. 14-3-3 proteins are essential for RAS/MAPK
cascade signaling during pseudohyphal development in S. cerevisiae. Cell 89:1055–65
93. Rottmann M, Dieter S, Brunner H, Rupp S. 2003. A screen in Saccharomyces cerevisiae
identified CaMCM1, an essential gene in Candida albicans crucial for morphogenesis. Mol.
Microbiol. 47:943–59
94. Russell C, Lay KM. 1973. Natural history of Candida species and yeasts in the oral cavities
of infants. Arch. Oral Biol. 18:957–62
95. Saiman L, Ludington E, Pfaller M, Rangel-Frausto S, Wiblin RT, et al. 2000. Risk factors
for candidemia in Neonatal Intensive Care Unit patients. The National Epidemiology of
Mycosis Survey study group. Pediatr. Infect. Dis. J. 19:319–24
Annu. Rev. Microbiol. 2005.59:113-33. Downloaded from arjournals.annualreviews.org

96. Sanchez-Martinez C, Perez-Martin J. 2002. Gpa2, a G-protein alpha subunit required

by Universidade Federal de Santa Catarina on 08/07/10. For personal use only.

for hyphal development in Candida albicans. Eukaryot. Cell 1:865–74

97. Savage DC, Dubos RJ. 1967. Localization of indigenous yeast in the murine stomach. J.
Bacteriol. 94:1811–16
98. Schembri MA, Kjaergaard K, Klemm P. 2003. Global gene expression in Escherichia coli
biofilms. Mol. Microbiol. 48:253–67
99. Scherwitz C. 1982. Ultrastructure of human cutaneous candidosis. J. Invest. Der-
This article
demonstrates the matol. 78:200–5
deformation of 100. Schinabeck MK, Long LA, Hossain MA, Chandra J, Mukherjee PK, et al. 2004. Rabbit
human corneocytes model of Candida albicans biofilm infection: liposomal amphotericin B antifungal lock
by C. albicans therapy. Antimicrob. Agents Chemother. 48:1727–32
hyphae during 101. Sherwood J, Gow NA, Gooday GW, Gregory DW, Marshall D. 1992. Contact sensing
hyphal penetration.
in Candida albicans: a possible aid to epithelial penetration. J. Med. Vet. Mycol. 30:461–69
102. Sherwood-Higham J, Zhu WY, Devine CA, Gooday GW, Gow NA, Gregory DW. 1994.
Helical growth of hyphae of Candida albicans. J. Med. Vet. Mycol. 32:437–45
103. Sohn K, Urban C, Brunner H, Rupp S. 2003. EFG1 is a major regulator of cell wall
dynamics in Candida albicans as revealed by DNA microarrays. Mol. Microbiol. 47:89–102
104. Soll DR, Galask R, Schmid J, Hanna C, Mac K, Morrow B. 1991. Genetic dissimilarity
of commensal strains of Candida spp. carried in different anatomical locations of the same
healthy women. J. Clin. Microbiol. 29:1702–10
105. Spoering AL, Lewis K. 2001. Biofilms and planktonic cells of Pseudomonas aeruginosa have
similar resistance to killing by antimicrobials. J. Bacteriol. 183:6746–51
106. Stanley NR, Lazazzera BA. 2004. Environmental signals and regulatory pathways that
influence biofilm formation. Mol. Microbiol. 52:917–24
107. Stoldt VR, Sonnenborn A, Leuker CE, Ernst JF. 1997. Efg1p, an essential regu-
The authors
identify EFG1 as a lator of morphogenesis of the human pathogen Candida albicans is a member of a
key regulator of conserved class of bHLH proteins regulating morphogenetic processes in fungi.
morphogenesis in EMBO J. 16:1982–91
C. albicans. 108. Suci PA, Tyler BJ. 2003. A method for discrimination of subpopulations of Candida albi-
cans biofilm cells that exhibit relative levels of phenotypic resistance to chlorhexidine. J.
Microbiol. Methods 53:313–25
109. Sweet SP. 1997. Selection and pathogenicity of Candida albicans in HIV infection. Oral
Dis. 3(Suppl. 1):S88–95
110. Tebarth B, Doedt T, Krishnamurthy S, Weide M, Monterola F, et al. 2003. Adaptation
of the Efg1p morphogenetic pathway in Candida albicans by negative autoregulation and
PKA-dependent repression of the EFG1 gene. J. Mol. Biol. 329:949–62

132 Kumamoto · Vinces

 ANRV253-MI59-06 ARI 4 August 2005 11:16

111. Vilain S, Cosette P, Zimmerlin I, Dupont JP, Junter GA, Jouenne T. 2004. Biofilm
proteome: homogeneity or versatility? J. Proteome Res. 3:132–36
112. Villar CC, Kashleva H, Dongari-Bagtzoglou A. 2004. Role of Candida albicans polymor-
phism in interactions with oral epithelial cells. Oral Microbiol. Immunol. 19:262–69
113. Watts HJ, Very A-A, Perera THS, Davies JM, Gow NAR. 1998. Thigmotropism and
The authors
stretch-activated channels in the pathogenic fungus Candida albicans. Microbiology demonstrate that
144:689–95 inhibition of MS
114. Weide MR, Ernst JF. 1999. Caco-2 monolayer as a model for transepithelial migration channels inhibits
of the fungal pathogen Candida albicans. Mycoses 42(Suppl. 2):61–67 thigmotropism in
C. albicans.
115. Wiesner SM, Bendel CM, Hess DJ, Erlandsen SL, Wells CL. 2002. Adherence of yeast
and filamentous forms of Candida albicans to cultured enterocytes. Crit. Care Med. 30:677–
83
116. Xiao JZ, Watanabe T, Kamakura T, Oshima A, Yamaguchi I. 1994. Studies on the cellu-
Annu. Rev. Microbiol. 2005.59:113-33. Downloaded from arjournals.annualreviews.org

lar differentiation of Magnaporthe grisea. Physiological aspects of substratum surfaces in

by Universidade Federal de Santa Catarina on 08/07/10. For personal use only.

relation to appressorium formation. Physiol. Mol. Plant Pathol. 44:227–36

117. Yamada-Okabe T, Mio T, Ono N, Kashima Y, Matsui M, et al. 1999. Roles of three
histidine kinase genes in hyphal development and virulence of the pathogenic fungus
Candida albicans. J. Bacteriol. 181:7243–47
118. Zhou XL, Kung C. 1992. A mechanosensitive ion channel in Schizosaccharomyces pombe.
EMBO J. 11:2869–75

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Annual Review of
Microbiology

Volume 59, 2005

Contents

Frontispiece
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Georges N. Cohen p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p xiv

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Looking Back
Georges N. Cohen p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p 1
Signaling in the Arbuscular Mycorrhizal Symbiosis
Maria J. Harrison p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p19
Interplay Between DNA Replication and Recombination in
Prokaryotes
Kenneth N. Kreuzer p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p43
Yersinia Outer Proteins: Role in Modulation of Host Cell Signaling
Responses and Pathogenesis
Gloria I. Viboud and James B. Bliska p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p69
Diversity and Evolution of Protein Translocation
Mechthild Pohlschröder, Enno Hartmann, Nicholas J. Hand, Kieran Dilks,
and Alex Haddad p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p91
Alternative Candida albicans Lifestyles: Growth on Surfaces
Carol A. Kumamoto and Marcelo D. Vinces p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p 113
Yeast Evolution and Comparative Genomics
Gianni Liti and Edward J. Louis p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p 135
Biology of Bacteriocyte-Associated Endosymbionts of Plant
Sap-Sucking Insects
Paul Baumann p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p 155
Genome Trees and the Nature of Genome Evolution
Berend Snel, Martijn A. Huynen, and Bas E. Dutilh p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p 191
Cellular Functions, Mechanism of Action, and Regulation of FtsH
Protease
Koreaki Ito and Yoshinori Akiyama p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p 211

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Mating in Candida albicans and the Search for a Sexual Cycle

R.J. Bennett and A.D. Johnson p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p 233
Applications of Autofluorescent Proteins for In Situ Studies in
Microbial Ecology
Estibaliz Larrainzar, Fergal O’Gara, and John P. Morrissey p p p p p p p p p p p p p p p p p p p p p p p p p p p p 257
The Genetics of the Persistent Infection and Demyelinating Disease
Caused by Theiler’s Virus
Michel Brahic, Jean-François Bureau, and Thomas Michiels p p p p p p p p p p p p p p p p p p p p p p p p p p p p p 279
Intracellular Compartmentation in Planctomycetes
John A. Fuerst p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p 299
Biogenesis of Inner Membrane Proteins in Escherichia coli
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Joen Luirink, Gunnar von Heijne, Edith Houben,

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and Jan-Willem de Gier p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p 329

Genome-Wide Responses to DNA-Damaging Agents
Rebecca C. Fry, Thomas J. Begley, and Leona D. Samson p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p 357
The Rcs Phosphorelay: A Complex Signal Transduction System
Nadim Majdalani and Susan Gottesman p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p 379
Translational Regulation of GCN4 and the General Amino Acid
Control of Yeast
Alan G. Hinnebusch p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p 407
Biogenesis, Architecture, and Function of Bacterial Type IV Secretion
Systems
Peter J. Christie, Krishnamohan Atmakuri, Vidhya Krishnamoorthy,
Simon Jakubowski, and Eric Cascales p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p 451
Regulation of Bacterial Gene Expression by Riboswitches
Wade C. Winkler and Ronald R. Breaker p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p 487
Opportunities for Genetic Investigation Afforded by Acinetobacter
baylyi, A Nutritionally Versatile Bacterial Species that is Highly
Competent for Natural Transformation
David M. Young, Donna Parke, and L. Nicholas Ornston p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p 519
The Origins of New Pandemic Viruses: The Acquisition of New Host
Ranges by Canine Parvovirus and Influenza A Viruses
Colin R. Parrish and Yoshihiro Kawaoka p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p 553
Vaccine-Derived Polioviruses and the Endgame Strategy for Global
Polio Eradication
Olen M. Kew, Roland W. Sutter, Esther M. de Gourville, Walter R. Dowdle,
and Mark A. Pallansch p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p 587

viii Contents
 Contents ARI 8 August 2005 15:56

INDEXES

Subject Index p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p 637

Cumulative Index of Contributing Authors, Volumes 55–59 p p p p p p p p p p p p p p p p p p p p p p p p p p p 675
Cumulative Index of Chapter Titles, Volumes 55–59 p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p p 678

ERRATA

An online log of corrections to Annual Review of Microbiology chapters

may be found at http://micro.annualreviews.org/
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Contents ix