Fungal Cells

Secondary article
Article Contents

Nawin C Mishra, University of South Carolina, Columbia, South Carolina, USA

. Introduction

Fungal cells have been used to produce a variety of useful items such as bread, beer, cheese
and antibiotics. The study of fungal cells has also facilitated advances in medicine and
agriculture.

. Cell Organization and Structure

. Nutrition
. Cellular Communication
. Yeast

Introduction
Fungi represent a myriad of cell types from unicellular
yeast to multicellular mushrooms with a very high level of
structural organization. Fungi have been utilized to
produce a variety of things used in daily life such as bread,
beer, cheese, wine, antibiotics and now possibly an insulinmimetic chemical to be developed as insulin pills for
diabetic patients. Fungi cause several human diseases from
athletes foot to meningitis. They usually attack immunocompromised individuals causing deadly infections. Most
AIDS patients suer from severe complications of fungal
infections. In plants, fungi cause havoc; the infamous
potato blight in the nineteenth century wiped out potatoes
and led to a great famine in Ireland. Fungi such as
mushrooms and morels provide valuable food for humans.
However, consumption of certain mushrooms can cause
food poisoning and people have died after eating mushrooms in the wild. Fungi, with a simple life style and genetic
organization similar to that of humans, have been utilized
to elucidate certain very important principles in the science
of genetics. Studies of fungi have indeed brought revolutions in genetics and molecular biology leading to an
understanding of the mechanism of gene action, feasibility
of gene cloning, and projects like the human genome
project and possibilities for gene therapy and gene designed
drugs.
A detailed understanding of the fungi has been instrumental in saving crops from fungal infections that could
otherwise wipe out up to 60% of present crop yield.
Understanding of fungi, as an organism at the cellular and
molecular levels, is crucial in developing strategies for
controlling human diseases caused by fungi.
Except for unicellular yeasts, fungi are lamentous. A
number of fungi like Candida can exist as unicellular yeast or
as lamentous fungus. Filamentous fungi are usually
multinucleate and dierentiate into distinct vegetative
forms such as aerial hyphae, conidiophores, conidia and
other asexual spores and structures for sexual reproduction.
Fungi also produce an array of structures in response to
environmental stimuli which initiate and establish infections, growth and other responses, such as circadian rhythm
and production of antibiotics or toxins. Certain fungi have
precise mating responses and many produce elaborate and
complex sexual fruiting structures leading to sexual spore
(such as ascospore and basidiospore) formation.

Cell Organization and Structure

Fungal cells like other eukaryotes possess at least three
reservoirs of genetic information located in the nucleus,
mitochondria and plasmids. The major genetic material is
organized into chromosomes and housed inside a nucleus
with nuclear membrane. The fungal cells divide either by
mitosis or meiosis. Like any other eukaryote, fungal cells
contain several organelles such as mitochondria, ribosomes, dictyosomes and nuclear membrane. Fungal cells
resemble plant cells and bacteria in having a thick cell wall
that is fused to the cell membrane, but dier from plant
cells by lacking a chloroplast, and from bacteria in the
organization of the genetic material. Bacteria are prokaryotes and lack nuclear organization and contain genetic
material directly in the cytoplasm. Fungal cells resemble
animal cells except for the thick cell wall. Animal cells lack
cell walls and the contents of cells are held inside only by a
cell membrane. Certain components of the fungal cell
membrane are distinct from the animal cell membrane,
which is the basis of certain antibiotic treatments to ght
fungal infection in humans and other organisms.

Organization of genetic material

Fungal cells contain one or more nuclei. Yeast cells are
typically uninucleate whereas the majority of lamentous
fungi are multinucleate. A number of lamentous fungi are
ceonocytic because the septum separating the adjacent cells
is not complete and therefore the nuclei readily move from
one part of a lament to another as a result of cytoplasmic
streaming. In ceonocytic fungi, the nuclear membrane
remains intact during the process of mitosis in order to
avoid the risk of mixing chromosomes among dierent
nuclei undergoing nuclear division at the same time.
Genetic control of the cell cycle and the separation of
chromosomes during cell division have been elucidated to a
great extent in yeast cells (Orr-Weaver, 1999). Ordinarily,
cohesin, a protein, keeps the sister chromatids held
together. However, as the cell division progresses toward
anaphase, separin, a protease, existing as an inactive
complex of separin and securin, is released due to the
digestion of securin by another protease called APC

ENCYCLOPEDIA OF LIFE SCIENCES 2001, John Wiley & Sons, Ltd. www.els.net

Fungal Cells

(anaphase promoting complex). Thus the released separin,

which becomes an active protease, degrades cohesin and
facilitates the sister chromatids separation. Yeast genes
Scc1, Esp1 and Pds1 encode cohesin, separin and securin,
respectively, and control the separation of sister chromatids during mitosis; homologues of these yeast genes have
been known to exist in vertebrates.

Nucleus
Ordinarily, fungal nuclei contain one set of chromosomes
and are haploid. However, certain fungi exist as diploid or
polyploid organisms. Yeast cells exist both as haploid and
diploid. All haploid fungi become diploid after fertilization
leading to the formation of a zygote, but this is a temporary
phase since the zygote undergoes meiosis to form four or
more (due to subsequent mitosis) haploid spores, which
may exist as unicellular or multicellular organisms. The
sexual phase, involving zygote formation and meiosis, has
not yet been identied in a number of fungi called fungi
imperfecti, which reproduce asexually. Among certain
ascomycetous fungi, the four products of meiosis, i.e.
ascopores, are retained in a bag-like structure called an
ascus. Analysis of ascospores has provided insight into the
role of a gene in controlling a particular phenotype and the
linkage relationship of that gene to the centromere and to
other genes on a chromosome.
The nuclear chromosome is typically a linear structure
with chromatids, centromere and telomeres; the chromatid
contains several origins of replication or ARS (autonomously replicating sequences) necessary for initiation of
replication. The mitochondrial chromosome is a circular
structure very much like the bacterial chromosome but
much smaller than the bacterial chromosome. Plasmid
DNA exist freely in the nucleus, mitochondria or
cytoplasm. They may integrate into nuclear or mitochondrial DNA.

Cell wall and membrane

The organization of fungal cells is that of a typical
eukaryotic cell. The cellular organization has been
visualized by both transmission and scanning electron
microscopy (EM). Such EM studies show the presence of a
complex cell wall fused with the cell membrane and the
presence of organelles including nucleus, mitochondria,
dictyosomes and ribosomes. Certain fungi, such as
Neurospora, contain deposits of ergesterol in the cytoplasm
as dense bodies. The cell membrane is a typical bilayer lipid
structure. Fungal cell membranes dier from animal and
other cell membranes in being sensitive to nystatin,
amphotericin B and ketoconazole which can punch holes
in the fungal cell membrane leading to lysis and breakdown
of the cell; this is the basis of treating patients with certain
fungal infections. Such drugs combine with sterol in the
2

fungal plasma membrane and lead to its disruption. These

antibiotics do not act on bacteria since the bacterial plasma
membrane lacks sterol. However, the plasma membrane of
animal cells does contain some sterol and therefore these
antibiotics could prove toxic to animal cells. Animal cell
membranes contain mostly cholesterol whereas fungal cell
membranes are made up of ergesterol, against which these
antibiotics are most eective; these dierences in the
chemical make-up of cell membranes render fungal cells
more readily sensitive to these antibiotics. Most morphological mutants are defective in the biosynthesis of the cell
wall. Also certain morphogenic substances such as lsorbose can alter the morphology of lamentous fungi and
make them grow as tight colonies on plates containing lsorbose. l-Sorbose inhibits the synthesis of a specic
polysaccharide, 1-3 b-glucan, in the cell wall. Certain
dimorphic fungi such as Candida and yeast which can grow
as unicellular or multicellular lamentous forms undergo
changes in cell wall biosynthesis in response to temperature
(in the case of Candida) or to nitrogen in growth medium
(in the case of yeast). As a defence mechanism against
fungi, certain plants upon fungal infection overproduce
chitinase to break down the cell wall of infecting fungus in
order to escape fungal attack. Chitin has been a major
target for developing antibiotics to control or eliminate
fungal infections. Fungi, like bacteria, are sensitive to sulfa
drugs since the latter interfere with the synthesis of the
precursor of folic acid; humans do not make folic acid but
obtain it from food; they are therefore not adversely
aected by sulfa drugs.

Genes and genomics

Genomic studies of a number of fungi of agricultural,
medical and industrial importance are underway (Goeau
et al., 1996; Kupfer et al., 1997). These include Saccharomyces cerevisiae (baking and brewing industries), Aspergillus nidulans and related Penicillium chrysogenum,
Trichoderma reesi (used for production of antibiotics and
several commercially important proteins) and Aspergillus
fumigatus and many other aspergilli (causing production of
toxin called aotoxin in post harvest crop). Candida
albicans and Cryptococcus neoformans, which cause infections of internal organs or meningitis in immunocompromised or AIDS patients, are also suciently important to
be included in such studies. These fungi have been
subjected to genomic analysis in order to study the
structure, function and evolution of fungal genes and
genomes. The genome of Saccharomyces cerevisiae is
completely sequenced. The size of fungal genomes varies
from 13.7 Mb to 250 Mb (Table 1). Typically the unicellular
yeast has a genomic size of 13.7 Mb with about 5800 genes,
suggesting a gene density of 470 genes/Mb, whereas the
multicellular fungi, such as Aspergillus, Neurospora and

Fungal Cells
Table 1 Genomics of certain fungi
Organism

aNo. of chromosomes
(linkage group)

Unicellular (yeast form)

Saccharomyces cerevisiae
Saccharomyces pombe

16
4

Multicellular (filamentous form)

Phytophthora infestans
Magnaporthe grisea
Aspergillus nidulans
Neurospora crassa

10

8
7

Total DNA
sequence (Mb)
13.5
14.0
250
47
31
42

Total No. of genes

estimated

Gene densityb

5800
7000

430
500

8500
8100
9200

225
240
225

Data based on molecular characterization of these organisms is by a number of investigators.

a
Chromosome members based on electrophoretic karyotyping as well as on cytogenetic analysis.
b
Gene/Mb of DNA segment.

others, possess a genome size of about 38 Mb with about

8000 genes and gene density of 230 genes/Mb.

Mitochondrial genome
In fungi, mitochondrial (mt) DNA is the second reservoir
of genetic information. Typically a fungal mitochondrion
contains circular DNA of about 65 kb, which can code for
about 15 proteins, several tRNAs and ribosomal RNAs. In
certain fungi, the mitochondria contain a linear DNA.
However, the size of mitochondrial DNA in fungi diers
markedly from that of animal mtDNA, the latter containing only about 15 kb of DNA and having almost the same
number of genes as fungal mtDNA. The size dierence
between the fungal mtDNA and the animal mtDNA is due
to the presence of G/C-rich spacers, certain tandem
duplications and introns in the fungal mtDNA, in addition
to a few extra genes such as mal gene required for metabolic
pathways unique to fungi.
Genes carried by mtDNA are for those proteins involved
in electron transport and oxidative phosphorylation that
are located in the mitochondrial membrane. A large
number of mitochondrial proteins are encoded by nuclear
genes which are translated on cytoplasmic ribosomes and
then transported into the mitochondria. Study of fungal
mitochondria has established the mode of organellar trac
of protein from cytoplasm to mitochondria and the
involvement of porin channels and chaperon proteins in
these processes. These proteins that are imported into
mitochondria possess an N-terminal signal sequence,
which interacts with receptors on the mitochondrial
surface requiring energy for their transport. Mitochondrial
DNA is uniparentally transmitted to the progeny of sexual
crosses. Fungal cells may harbour mitochondria with
complete full size mtDNA, and also mitochondria with
deleted mtDNA. In certain mutant strains, mtDNA with

deletions generates a minimal size mtDNA by tandem

duplication of the remaining DNA sequence. This shows
that mtDNA must attain a minimal size in order to exist.
Several human diseases also contain a mixture of
mitochrondria with full size mtDNA and mtDNA with
deletions. Study of the fungal mitochondria also established the departure from the universality of the genetic
code.

Plasmid genome
Plasmids represent the third reservoir of genetic information in fungi; they are 39 kb in length and occur as circular
or linear structures existing in the nucleus or in the
mitochondria. Of these the most thoroughly known
plasmid is the 2 mm circle (2mCi) of yeast that exists in the
nucleus. This is a circular plasmid with only a few genes, the
most important being the FLP gene encoding a recombinase. This is now exploited as a tool to cause site-specic
recombinations in dierent organisms. The 2mCi is an
episomal plasmid since it can exist autonomously or
integrate into yeast chromosome. Neurospora plasmids
are mitochondrial plasmids. They exist freely as circular or
linear DNA molecules or may integrate into mtDNA,
causing disruption of the mitochondrial genes and their
functions leading to senescence of fungus. The yeast 2mCi
plasmid defective in DNA replication has been used to
isolate and characterize small yeast chromosomal DNA
segments which can endow an ability for DNA replication.
These DNA segments are called ARS (autonomously
replicating sequences). Addition of these DNA sequences
can restore the ability of a replication-defective plasmid to
replicate. Identication and characterization of such ARS
has been instrumental in the construction of yeast articial
chromosomes as described later. This yeast plasmid has
been extensively used as a vector for gene transfer in yeast.
3

Fungal Cells

The lamentous fungal plasmids are being redesigned to be

utilized as a vector for gene transfer and used as a vehicle
for the transfer of genes in other fungi.

Nutrition
The minimal nutritional requirements of fungi, and the fact
that a recipe for such growth medium was established as
early as the 1930s, have been the major factors in the study
of fungi under laboratory conditions for genetic and
biochemical analyses. Such biochemical genetic approaches were responsible for establishing metabolic
pathways and their genetic control leading to the formulation of the one geneone enzyme hypothesis by Beadle and
Tatum in 1941. The impact of the one geneone enzyme
hypothesis is such that one must study a mutant rst via
genetic dissections or molecular methods in order to
establish the characteristics of the normal physiology of
any organism from viruses to humans.
An understanding of the nutrition of fungi was also
crucial in developing the methodology for large-scale
preparation of fungal proteins, enzymes and secondary
metabolites including antibiotics and mycotoxins. The
secondary metabolites are those substances that fungi
synthesize for their survival in a particular environment;
however, these substances do not play any role in the
growth of the producing fungus. A fungus commits a large
proportion of its cellular energy to producing these
substances in order to ensure its existence in a particular
ecological niche. The secondary metabolites are of
immense signicance to humans and have been harnessed
by developing biotechnology to produce them on a larger
scale. One of the major secondary metabolites is antibiotics
such as penicillin. The pathway for penicillin biosynthesis
is almost fully understood and even the genes encoding the
proteins for producing antibiotics have been cloned and
harnessed to some extent. The other secondary metabolite
that has been much investigated is aatoxin, a polyketide
secondary metabolite. This mycotoxin is believed to cause
cancer of the liver in humans and several disorders in
animals. Aatoxicosis results from the consumption of
grains with post-harvest growth of fungi, such as
Aspergillus avus, A. nidulans and A. parasiticus. The
biochemical pathway involving more than 10 steps has
been fully characterized. In addition, the gene involved in
the regulation of aatoxin biosynthesis has been identied
and characterized. The regulatory gene ar encodes a
DNA-binding protein belonging to the class of regulatory
proteins such as Gal4 protein in yeast. A large number of
fungi establish symbiotic associations with plants and
obtain carbon from plants while providing them with
nitrogen. The genetic control of such nutritional exchange
and the biochemical mechanism of mycorrhizal formation
are not yet fully understood.
4

Cellular Communication
Intercellular communication in fungi is known to facilitate
such complex functions as cell cycle, mating, hyphal
dierentiation, infections and heterokaryosis. In addition,
signal transduction or intracellular communication is used
for regulation of gene action and control of vital functions
like circadian rhythm. In yeast, the molecular biology of
signal transduction is elucidated to a great extent via the
dissection of appropriate genetic mutants. It is known to
involve a cascade of genes, which in response to a specic
external stimulus, control a number of mitogen-activated
protein kinases (MAPK), which in turn control genes for
specic transcription factors leading to transcription of
genes involved in specic metabolic pathways. In yeast at
least six MAPK pathways are established that respond to
pheromones, nutritional substances or starvation, protein
kinase C, RAS protein and other stimuli, leading to a
number of developmental pathways including mating,
lamentous growth, osmolarity-response, infectivity and
other responses. Genes and proteins involved in dierent
signal transduction pathways of yeast have been characterized. Some of these signal transduction pathways are
also found to be common to other fungi including
Neurospora, Ustilago and Candida.

Mating response in fungi

Fungi are usually heterothallic (self-sterile) or homothallic
(self-fertile) or pseudohomothallic. In heterothallic fungi,
the gametes come from dierent parents and are usually
controlled by a dierence in one mating type locus, as in
Neurospora crassa, or by many alleles, as in Schizosaccharomyces commune. Homothallic fungi involve the fusion of
gametes with identical nuclei and are not controlled by a
mating type locus. Pseudohomothallic fungi, although
self-fertile, involve gametes with nonidentical nuclei such
as the case in Neurospora tetrasperma. The mating type loci
in fungi (as exemplied by Neurospora and other fungi) do
not qualify as alleles or allomorphs in a true sense; instead,
they represent idiomorphs. The chromosomal segments
controlling the alternative mating type response occupy
the same position on the chromosome but dier in their
nucleotide sequence; therefore they are idiomorphs. In
Neurospora crassa, the two mating type loci A and a,
identied as 5.3 and 3.2 kb DNA segments, respectively,
located on chromosome I, control the mating type
reactions which include fusion of trichogyne to conidium,
secretions and response to pheromones, perithecial development and ascospore formation. The mating type A locus
comprises 5301 bp; of this only a 750 bp segment controls
the mating response. This contains an open reading frame
(ORF) capable of encoding a protein of 228 amino acids
similar to yeast mating type MAT alpha-1. The other
Neurospora mating type, a, consists of 3235 bp, of which

Fungal Cells

only a 1260 bp segment controls the mating type response.

This segment encodes a protein of 382 amino acids which is
similar to mat-Mc-encoded protein in Saccharomyces
pombe. The N-terminus and C-terminus of this 382-amino
acid protein controls the mating type reaction and the
vegetative incompatibility, respectively.
The opposite mating type strains of all heterothallic
species (such as Neurospora crassa, N. sitophila, N.
intermidia and N. discreta) contain either the A or a
DNA segment. In the pseudo-homothallic species such as
N. tetrasperma, self-fertile cultures are heterokaryotic with
respect to mating type nuclei A and a and fusion leading to
mating type reaction involves these nuclei with opposite
mating types A and a. In contrast, the homothallic species
such as N. africana and N. dodgi contain only A idiomorph.
However, N. terricola possesses both A and a idiomorphs.
In homothallic species it seems that the target gene for the
mating type idiomorph responds only to A idiomorph. The
a mating type idiomorph has undergone mutation in the
course of evolution and it has lost its activity. A large
number of other ascomycetous fungi have the two mating
type idiomorphs as elucidated in N. crassa. Among
basidiomycetes sexuality is controlled by either one or
two pairs of alleles or idiomorphs. These respectively
represent bipolar and tetrapolar systems. In both unicellular yeast and in a number of lamentous fungi, there is
a mating type switch leading to homothallic (self-fertile)
pattern. In yeast the gene responsible for such mating type
switch called HO (for homothallism) has been very well
characterized. The HO gene of lamentous fungi has not
yet been characterized at the molecular level. Neurospora
diers from yeast in that it has no extra copy of mating type
idiomorph; therefore, mating type switch is not possible in
Neurospora. Several asexual fungi carry a mating type
locus, which is functional when transferred to a sexual
fungus with a defective mating type locus, but is not
functional in the resident asexual fungus. This suggests
that mating type locus alone does not control sexual
response in fungi.
Mating type response is genetically controlled by one or
more pairs of genes; their gene products control a signal
transduction pathway and its regulation. Mating response
involves participation by hydrophobic peptide pheromones produced in sex-specic mating, secretion of
these pheromones and their processing. These pheromones
are perceived by receptors found as transmembrane
proteins which are linked to protein-kinase signal transduction pathways via hetrotrimeric G proteins. The
interactions between opposite mating type strains lead to
growth response, spatial dierentiation, activation of
responsive genes and nally fusion of cells by mating type
agglutinins culminating into cell fusion and then nuclear
fusion and meiosis with simultaneous elaboration of the
fruiting body.

Cell fusion and heterokaryosis

A large number of fungi undergo cell fusion or hyphal
fusion, thereby causing heterokaryosis, i.e. existence of two
kinds of nuclei in the common cytoplasm. Heterokaryosis
is a prelude to sexual reproduction in a number of
lamentous fungi, particularly among the basidiomycetes.
Such fusion of vegetative hyphae leading to heterokaryosis
provided geneticists with the opportunity for determination of the dominance and recessiveness of dierent alleles
of a gene and for complementation analysis of mutants
leading to their identication as mutations occurring in the
same gene or in dierent genes. Later such complementation became the basis for identication of the function of a
cloned DNA segment by its ability to rescue a mutant
strain.
In Neurospora such cell-to-cell interaction has been
found to be genetically controlled. Besides the idiomorph
of the mating type locus, at least 10 other genes are known
to control compatibility for heterokaryosis in Neurospora.
Strains must be isogenic for the Het genes in order to
facilitate cell fusion leading to heterokaryon formation.
Non-isogenic strains are vegetatively incompatible and
cannot form a heterokaryon. The vegetative incompatibility is manifested in several ways including lack of
continued growth after stable heterokaryon formation.

Hyphal differentiation
Morphogenesis has been extensively studied in yeast and
Neurospora. It seems signal transduction plays an important role in the development of the hyphae. The
environmental signals are transduced by heteromeric G
proteins to receptors in the plasma membrane. Neurospora
mutants defective in heteromeric G protein have lower
levels of intracellular cAMP and are also defective in
hyphal and conidial development. They show increased
female fertility in the form of enlarged perithecia and
express more sensitivity to heat and oxidative stress. Thus
it seems that a G alpha protein mediates signal transduction pathways in Neurospora via control of cAMP levels
leading to hyphal dierentiation and modulations of stress
response. The process of hyphal dierentiation is also
critical for dimorphic changes in several fungi and the
infectivity of pathogenic fungi. In yeast hyphal dierentiation is almost fully established (Liu et al., 1993; Fujita et al.,
1999; Herskowitz, 1995). This involves a cascade of signal
transduction in response to nitrogen starvation involving a
number of genes leading to pseudohyphal growth. In yeast
these include genes for protein kinase (Ste20p), MAPKKK
(Ste11p), MAPKK (Ste7p), MAPK (Fu3p) and MAPK
(Ksslp). The Ste20p gene controls genes for MAPK and
MAPKK and leads to pseudohyphal development in
response to nitrogen deciency. Another gene, Hs17p, acts
as a negative regulator of Ste20p such that a hs17p mutant
can lead to pseudohyphal development even in the
5

Fungal Cells

nitrogen-rich medium. The role of RAS proteins has been

found to be common in all fungi undergoing hyphal
development.

Conidiation, circadian rhythm and

biological clock
Conidiation in lamentous fungi is a major event of
dierentiation. During conidiation, entirely dierent sets
of genes are expressed as evidenced by the new prole of
mRNA produced during conidiation. Conidiation in
certain lamentous fungi such as Neurospora shows some
features of circadian rhythm. In addition to conidiation,
many other physiological activities are controlled by
circadian rhythms. Circadian rhythm is the manifestation
of a biological clock in all organisms (Bell-Pederson, 1998;
Dunlap, 1999). The components of this biological clock are
genetically determined. In Neurospora, it seems that blue
light is perceived by certain avonoid receptors and in
response to this, two transcription factor genes, WC-1 and
WC-2, are induced, products of these genes control the
transcription of freq-1 gene encoding another transcription
factor. The freq-1 gene is translated from two dierent
initiation codons ATG, located several nucleotides apart in
the mRNA, and produces proteins of dierent lengths;
these move into the nucleus and work as a feedback loop to
control the transcription of many other genes involved in
the rhythmic control of dierent physiological functions
including conidiation. In addition to WC-1, WC-2 and
freq-1 genes encoding dierent transcription factors, there
are a number of other genes that regulate the clock.
Proteins with similar domains and functions analogous to
the proteins encoded by Neurospora WC-1, WC-2 and freq1 genes have been identied in Drosophila and mouse. Thus
it seems that the components of the biological clock have
been conserved throughout the living system during the
process of evolution in order to coordinate dierent
physiological functions.

Infectivity
Infectivity by a fungal pathogen involves an array of signal
transduction on the part of pathogen and host. Infectivity
is usually associated with the ability to grow or undergo
switch from unicellular to lamentous forms of growth, to
adhere to various host surfaces and the production and
secretion of hydrolytic enzymes such as proteinases. All
these aspects involve genetically controlled signal transduction. Pathogen and host have evolved parallel genetic
systems which enable the pathogen to initiate an attack on
the host and also enable the host to resist attack by the
pathogen. Such response between host and pathogen is
called gene-for-gene type response. The pathogens usually
make proteinases whereas the host elaborates on specic
proteinase inhibitors. The product of Avr gene in the
6

pathogen controls proteinase production, whereas the

product of R gene controls the host response. Products of
both pathogen Avr gene and host R gene are essentially
transcription factors with dierent DNA-binding motif or
domain for proteinprotein interactions or proteinligand
interaction.
The earlier stages in infectivity by pathogenic fungus are
controlled by genes responsible for growth or dimorphic
dierentiation. These stages are genetically controlled by
the fungal cells based on the cue or signals that they get
from the host cell surface. For example, SAP-2 gene
functions are involved in the initial stage of infection by
Candida albicans and are essential for infection into deep
organs. Fungal infectivity may be acquired by horizontal
transmission without involving sexual mating even though
the sexual and asexual fungi may dier in their degree of
infectivity.

Signal transduction involved in

gene regulation
In fungi, the regulation of metabolism involving amino
acids, nitrogen, sulfur, phosphate and dierent carbon
sources is mediated by signal transduction leading to
activation of genes encoding dierent transcription factors
in response to environmental stimuli. The control of amino
acid synthesis was rst described in Neurospora; later its
molecular aspect was elucidated in yeast as GCN4 protein
and in Neurospora as CPC-1 gene product. Both yeast
GCN4 and Neurospora CPC-1 gene products promote the
transcription of genes involved in amino acid synthesis by
binding to TGACTC located in the 5 nontranscribed
regions of these genes. Both GCNA4 and CPC-1 proteins
have DNA-binding domains of the V-jun-encoded protein
and other leucine zipper proteins. Likewise other regulatory metabolic pathways involving sulfur, phosphate and
unusual carbon sources such as quinic acid have been
established.

Yeast
Study of Neurospora, Aspergillus and yeast has been
instrumental in the understanding of the various aspects
of eukaryotic genetics. The success of these studies has
culminated in the understanding of gene action (the one
geneone enzyme hypothesis of Beadle and Tatum, 1941)
and control of metabolic pathways; genetic mapping,
nature of recombination and gene conversion and their
underlying mechanism via formation and resolution of
heteroduplex or hybrid DNA called Holliday structure.
Understanding of parasexual recombination in Aspergillus
paved the way for analysis of linkage (or synteny) in
humans (via fusion of somatic cells). Development of a
gene transfer system in fungi (Mishra and Tatum, 1973),

Fungal Cells

particularly in yeast (Hinnen et al., 1978), marked the

beginning of the bonanza in the molecular biology of
eukaryotes that led to the understanding of almost all
features of an organism at the molecular level including the
DNA sequencing of the whole genome of yeast. Molecular
biology of yeast has led to the development of new
branches of genetics or related sciences such as genomics,
proteomics and bioinformatics.
The methodology for gene disruptions in yeast led
conceptually to development of knockouts in mice which is
so crucial for the understanding of the essential functional
role of a gene in the life of a mouse. Likewise, site-specic
recombination has been useful in control of gene activation. Development of gene transfer in yeast made it
possible to study the genetics of many other fungi,
particularly the asexual fungi, such as Candida albicans,
which is not amenable to classical mendelian analysis.
Genetic analyses of these organisms have been complemented by molecular and electrophoretic karyotyping rst
developed in yeast. Thus a large number of fungi have
become amenable to genetic analysis via gene transfer and
electrophoretic karyotyping. Yeast has also provided
insight into the understanding of dierent aspects of cell
growth and molecular interactions in gene control and
signal transduction via isolation and characterization of
temperature-sensitive (i.e. conditional) mutants defective
in various stages of the cell cycle. Development of the yeast
two-hybrid system has been crucial in identifying the
essential components of the signal transduction network.
This is accomplished by identication of unknown partner
protein able to restore normal interactions with a known
component protein leading to facilitation of proper
transcriptional control of a function in yeast. Thus the
yeast two-hybrid system has become a useful tool for
deciphering the nature of the multi-components of a
biological regulatory system and in establishing the
biological components of a regulatory control system
from evolutionarily diverse systems by their ability for
interaction leading to restoration of a function in yeast.
The yeast gene transfer system has also been used to
identify and characterize the dierent components of a
typical eukaryotic chromosome. These include ARS
elements (i.e. DNA segments that are origins of replication), responsible for autonomous replication of a
chromosomal segment; centromere, responsible for proper
distribution of chromosomes during cell division and
crucial for preserving the constancy of chromosome
numbers in the cells of a species; and telomeres (i.e. the
ends of a chromosome), responsible for maintaining the
integrity of a chromosome. Yeast articial chromosome
(YAC) can be created by proper assembly of these
components. Successful construction of YAC also requires
additional lengths of any DNA segment to attain a
minimum size. If YAC is less than 150 kb in length it is
not properly maintained during the cell cycle of yeast. This
requirement has been exploited to include up to 1 Mb of

alien chromosomal segment to create an array of YACcarrying overlapping alien DNA segments. Such an array
of YAC, called contigs, facilitated the linkage analysis and
nally the nucleotide sequencing of humans and other
organisms. Thus YAC has been a very crucial development
for the human genome project. Construction of YAC has
led to development of BAC (bacterial articial chromosome). Recently BAC has also been extensively used in
genomic analysis.
The development of YAC has led nally to the
construction of human articial chromosome (HAC).
HAC will be useful in gene therapy and will provide a
vehicle for the placement of a set of new genes in humans as
an additional chromosome. In the early part of the twentyrst century it is expected that humans may carry an
additional pair of HAC, making the total number of
chromosomes in the human cell 48 instead of the naturally
occurring 46 chromosomes. The additional chromosomes
may carry genes to overcome the deciency of defective or
missing genes and/or genes for enhancement of desirable
phenotypes such as higher intelligence or for controlled
expression of a desired gene at a desired time in the life of an
individual. All these developments will give rise to many
ethical concerns and considerations, requiring legal provisions in some areas.

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Fungal Cells

Further Reading
Allen MF (1992) Mycorrhizal Functioning: An Integrative Plant-fungal
Process. New York: Chapman and Hall.
Henning KA, Novotny EA, Compton ST et al. (1999) Human articial
chromosomes generated by modication of a yeast articial chromosome. Proceedings of the National Academy of Sciences of the USA 96:
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Mishra NC (1985) Gene transfer in fungi. Advances in Genetics 23: 73
177.

Mishra NC (1991) Genetics and molecular biology of Neurospora crassa.

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Mishra NC (1995) Molecular Biology of Nucleases. Boca Raton, FL:
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