Review
Initiation of Infection
Subversion of trafficking, apoptosis,
and innate immunity by type III
secretion system effectors
Benoit Raymond, Joanna C. Young, Mitchell Pallett, Robert G. Endres,
Abigail Clements, and Gad Frankel
Department of Life Sciences, Imperial College, London, UK
Injection of effector proteins by a type III secretion sys-
tem (T3SS) is a common infection strategy employed by
many important human pathogens, including enteric
Escherichia coli, Salmonella, Yersinia, and Shigella, to
subvert cell signaling and host responses. In recent
years, great advances have been made in understanding
how the T3SS effectors function and execute the diverse
infection strategies employed by these pathogens. In
this review, we focus on effectors that subvert signaling
pathways that impact on endosomal trafficking, cell
survival, and innate immunity, particularly phagocyto-
sis, nuclear factor-kB (NF-kB), and mitogen-activated
protein (MAP) kinase pathways and the inflammasome.
Type III secretion and infection
The intestinal epithelium provides a critical interface
between the body and the external environment acting as
a protective barrier against ingested toxins and patho-
gens and consists of the largest reservoir of microorgan-
isms that live in the body. Many pathogenic bacteria
have acquired sophisticated strategies to breach the
intestinal integrity and to exploit the mucosa as a niche
to replicate and disseminate into deeper tissues to cause
disease. Examples include enteropathogenic and enter-
ohemorrhagic Escherichia coli (EPEC and EHEC), Shi-
gella, Salmonella, and Yersinia species which all use
their type III secretion system (T3SS) to inject bacterial
effector proteins to subvert the host defenses and pro-
mote gut infection. Although these species all rely on
their T3SS for pathogenesis, their infection strategies
differ considerably. Whereas EPEC/EHEC and Yersinia
are predominantly extracellular pathogens, Salmonella
and Shigella trigger their own internalization and sur-
vive intracellularly. All of these pathogens use T3SS
effectors to regulate actin dynamics to facilitate their
own attachment or invasion, subvert endocytic traffick-
ing, block phagocytosis, modulate apoptotic pathways,
and manipulate innate immunity and host responses as
Corresponding author: Frankel, G. (g.frankel@imperial.ac.uk).
Keywords: enteropathogens; T3SS; subversion of cellular trafficking; phagocytosis
remodeling; cell survival modulation; inflammatory response manipulation.
0966-842X/$ – see front matter
ß 2013 Elsevier Ltd. All rights reserved. http://dx.doi.org/10.1016/j.tim.2013.06.008
430 Trends in Microbiology August 2013, Vol. 21, No. 8
part of the initial infection process and will be the focus
of this review. Although the actin cytoskeleton is manip-
ulated by T3SS effectors during infection, this has been
extensively reviewed elsewhere and thus will not be
covered in this review [1].
Disrupting cellular trafficking
Eukaryotic cells have two vesicular transport systems to
mobilize proteins and lipids. The secretory pathway trans-
ports material from the endoplasmic reticulum (ER) to the
plasma membrane (PM), via the Golgi, whereas the endo-
cytic pathway transports PM-derived endosomes to the
Golgi, ER, lysosomes (for degradation), or back to the
PM via recycling pathways. Both intracellular and extra-
cellular pathogens manipulate endocytic pathways using
T3SS effectors (Figure 1).
Secretory pathway
Transport steps in the secretory pathway involve cargo
packaging at the donor membrane followed by vesicle
budding, transport, tethering, and uncoating at the recep-
tor membrane. These processes are regulated by small
GTPases. Vesicles responsible for anterograde ER–Golgi
transport contain coat protein complex II (COPII), whereas
COPI coated vesicles are responsible for retrograde intra-
Golgi and Golgi–ER transport [2]. EPEC/EHEC translo-
cate several effectors which inhibit the secretory pathway,
including NleA/EspI, NleF, and EspG. NleA/EspI inhibits
the anterograde ER–Golgi transport by binding to Sec24
paralogs [3,4] of the COPII vesicle coat. NleA/EspI appears
to stabilize the COPII vesicles at the Golgi, inhibiting the
uncoating of the vesicle (with reduced hydrolysis of the
small GTPase Sar1) and hence reduced recycling of COPII
components back to the ER, resulting in reduced secretion.
The consequence of this reduced secretion during infection
is unclear, although NleA/EspI does significantly contrib-
ute to virulence of the EPEC/EHEC-like mouse pathogen
Citrobacter rodentium in vivo [5]. In polarized cultured
cells NleA/EspI can alter cellular tight junctions [6], po-
tentially by reducing secretion of replacement tight junc-
tion proteins. Recently, the EHEC effector NleF was shown
to bind Tmp21, a COPI vesicle receptor involved in trans-
Golgi network function, and may slow ER–Golgi and
 Review Trends in Microbiology August 2013, Vol. 21, No. 8

EPEC
EHEC

Salmonella Shigella

IpgD
PIP2 PIP
SopB Exo70
SipC

Lysosome

MPR Hydrolase
Hydrolase
MVB/ Exocyst
LE SKIP
SifA
SopB
MPR Salmonella Sif
SseF/G

IpaJ

ARF1
Golgi ARFs Rabs
EspG
Espl/NleA Tmp21 GM130 PAKs
Sec24 NleF

ER

Nucleus

TRENDS in Microbiology

Figure 1. Intracellular trafficking disruption. The EspI/NleA effectors from EPEC/EHEC (in blue) bind Sec24 to disrupt protein secretion, potentially by reducing recycling of
COPII to the ER. EspG acts as a Rab-GAP, but also interacts with ARFs, PAKs, and GM130 to disrupt protein secretion and NleF binds to Tmp21 to slow ER–golgi trafficking.
Shigella IpaJ (in orange) acts as cysteine protease that cleaves the N-myristoylated glycine from ARF1 to block cargo transport through the Golgi apparatus. SopB (in green)
and IpgD (in orange) from Salmonella and Shigella, respectively, dephosphorylate phosphatidylinositol 4,5 bisphosphate, PI(4,5)P2 to produce PI5P to regulate membrane
trafficking at the invasion site. SopB can also be found on the SCV. Salmonella SipC binds Exo70 to redirect exocysts to the bacterial attachment site. SseG and SseF
facilitate the extension of Sifs along the microtubules towards the cell periphery. Binding of SifA to SKIP sequesters Rab9, inhibiting the trafficking of MPR to the trans-Golgi
network and downstream trafficking of hydrolases to the lysosome and then the SCV. Abbreviations: EPEC/EHEC, enteropathogenic and enterohemorrhagic Escherichia
coli; COPII, coat protein complex II; ER, endoplasmic reticulum; Rab-GAP, Rab GTPase-activating protein; ARFs, ADP ribosylation factors; PAKs, p21 activated kinases; SCV,
Salmonella-containing vacuole; Sifs, Salmonella induced filaments; MPR, mannose phosphate receptor.

Golgi–PM trafficking, although further work is required to
clarify this function [7]. An additional EPEC/EHEC effec-
tor, EspG, was shown by multiple methods to disrupt the
secretory pathway [8,9]. EspG can function as a Rab
GTPase-activating protein (Rab-GAP) [10], trapping
Rab-GTPases in their inactive GDP bound form, and re-
ducing ER–Golgi transport. EspG can also bind numerous
other proteins including ADP ribosylation factor (ARF)
GTPases [8], p21 activated kinases (PAKs) [8,11], and
the Golgi matrix protein GM130 [9]. EspG interacts with
activated, membrane-embedded ARFs, potentially trans-
forming ARF activation into Rab inactivation at appropri-
ate endomembranes. During infection of cultured cells
EspG expression reduced interleukin-8 (IL-8) secretion
[10], suggesting it plays a role in immunomodulation.
The ability of NleA/EspI and NleF to reduce cytokine
expression during infection has not been reported, thus
it remains to be seen whether interrupting the secretory
pathway is another mechanism utilized by these patho-
gens to reduce immune recognition.
VirA, a homolog of EspG found in Shigella also acts as a
Rab-GAP but during Shigella infection it reduces autop-
hagic recognition (LC3 recruitment) of cytosolic Shigella,
thus protecting it from degradation [10]. Recently, the
Shigella effector IpaJ, a cysteine protease, has been shown
to cleave the myristoyl group from ARF1, which regulates
cargo transport through the Golgi apparatus, and is now
the second Shigella T3SS effector shown to alter the host
secretory pathway during infection [12].
In Salmonella, the C terminus of SipC interacts with the
exocyst targeting component Exo70 [13], causing exocysts to
tether at the bacterial attachment site, potentially provid-
ing extra membrane to facilitate invasion. To date, this is the
only Salmonella effector which specifically regulates the
host secretory pathway, although many Salmonella effec-
tors are involved in regulating endolysosome trafficking.
Endolysosome trafficking
The endosomal pathway internalizes extracellular materi-
al as well as plasma membrane proteins and lipids which
431
Review
are then sorted in the early endosome for degradation
through the late endosomes and lysosomes, or for transport
to the trans-Golgi, or back to the plasma membrane via
recycling endosomes. Once internalized by host cells Sal-
monella subverts the endolysosome trafficking pathway to
create a Salmonella-containing vacuole (SCV), which is
essential for its intracellular survival. The SCV transiently
acquires proteins from sorting and recycling endosomes
[Rab5, EEA1, and the transferrin receptor (TfR)] before
acquiring vacuolar ATPase, responsible for phagosome
acidification [14], and lysosomal glycoproteins (lgps), such
as Lamp1, in a Rab7-dependent process [15]. The SCV does
not however accumulate the lysosomal hydrolases or the
mannose phosphate receptors (MPRs) that traffic the
hydrolases from the Golgi to the lysosomes. This selective
exclusion has been attributed to the SPI-2 T3SS effector
SifA and its binding partner SKIP, which sequester Rab9
to inhibit MPR trafficking from late endosomes to the
trans-Golgi network and hence MPR-dependent trafficking
of the hydrolases from the Golgi to the lysosome [16]. As the
SCVs mature they travel to the microtubule organizing
center (MTOC), a perinuclear region adjacent to the Golgi,
where they produce Salmonella induced filaments (Sifs)
dependent on the SPI-2 T3SS effectors SseF and SseG. Sifs
extend along microtubules from the SCV to the cell periph-
ery where they recruit Rab9 and Rab11 and take up fluid
phase endocytic markers, potentially providing nutrients
for the bacteria from outside the cell [17–19].
By contrast, internalized Shigella avoids the endolyso-
some system by rapidly lysing its vacuole through the
action of the T3SS effectors IpaB and C [20]. IpaB in
particular has been shown to form membrane-disrupting
pores [20,21] and some studies suggest that IpaH7.8 may
also be involved [22]. Time-lapse analysis reported the
recruitment of host factors, including RhoA and Rac1, to
lysing vacuoles suggesting the involvement of host signal-
ing [23]; however, further research is required to fully
understand this key step in the pathogenesis of Shigella.
Phosphoinositide (PI) lipids play a significant regulato-
ry role in membrane trafficking. Salmonella SopB/SigD
and Shigella IpgD are PI phosphatases that dephosphory-
late phosphatidylinositol 4,5 bisphosphate, PI(4,5)P2, to
produce PI5P. Although SopB aids bacterial invasion by
localizing to the plasma membrane early in infection [24],
it later relocalizes to the SCV [25] where it alters the PI
composition to reduce the recruitment of Rab8, Rab13,
Rab23, and Rab35 and prevent phagolysosome formation.
The formation of PI5P by IpgD at the PM recruits epider-
mal growth factor receptor (EGFR) to the PM on early
endosomes to stop maturation and degradation leading to
prolonged cell survival through Akt activation [26]. Similar
to SopB, IpgD has pleiotropic effects, including cytoskele-
tal reorganization at the bacterial invasion site [27] and
cell migration, in addition to its role in cell survival.
Phagocytosis remodeling
Professional phagocytes of the immune system (including
neutrophils, macrophages, and dendritic cells) mediate the
ingestion of foreign material and pathogenic organisms,
which results in both destruction of the organism and
generation of peptides for MHC presentation to prime
432
Trends in Microbiology August 2013, Vol. 21, No. 8
the adaptive immune response. Phagocytes can recognize
bacteria directly [e.g., through scavenger receptor A recog-
nition of lipopolysaccharide (LPS) [28]] or indirectly
through recognition of immune opsonins (IgG and the
complement cascade component C3bi), which coat the
bacteria and are recognized by the Fcg receptors and
integrin amb2 (complement receptor 3), respectively
[29,30]. Receptor ligation activates complex signaling
pathways involving tyrosine kinases, small Rho GTPases
and PIs. Successful pathogens display an extensive array
of strategies for subverting phagocytosis (Figure 2). Intra-
cellular Salmonella and Shigella escape phagocytic killing
through manipulation of endolysosomal trafficking, as
discussed above, which disrupts phagosome maturation.
Yersinia cross the intestinal epithelial layer by trans-
cytosis through antigen sampling M cells and can then
invade macrophages and enterocytes mediated by an in-
teraction between Invasin and b1 integrins [31]. However,
Yersinia predominantly survives extracellularly using the
translocation of effectors to inhibit cytoskeletal rearrange-
ments and phagocytosis. With the exception of EspT-posi-
tive invasive strains (2% EPEC strains) [32], EPEC/
EHEC inhibit both transcytosis through M cells and inter-
nalization by phagocytes [33,34].
Yersinia inhibits phagocytosis through the concerted
actions of several effector proteins. YopH is a tyrosine
phosphatase, which acts on several cytoskeletal proteins
including p130Cas, Fyn binding protein (Fyb), focal adhe-
sion kinase (FAK), p85, Lyn, and paxillin [35–38]. Dephos-
phorylation of p130Cas disrupts its interaction with Crk
and subsequent Rac activation required for Yersinia inter-
nalization [39]. Yersinia also subverts the actin polymeri-
zation required for phagocytosis by the actions of several
effectors which manipulate Rho GTPases. The effector
YopE acts as a RhoGAP [40] and inhibits Rac-dependent
actin polymerization either directly or through inactiva-
tion of the upstream RhoG [41,42]. The effector YopO/YpkA
has a carboxy-terminal GDP-dissociation inhibitor (GDI)-
like domain which interacts with Rho GTPases [43] and is
suggested to sequester the GDI-free pool of Rac1 localized
at the membrane [44]. YopT, restricted to Yersinia enter-
ocolitica, acts as a cysteine protease cleaving the carboxy-
terminal prenylyl groups of RhoA, Rac1, and Cdc42 [45].
This cleavage inactivates Rho GTPases by disrupting their
membrane localization.
Extracellular EPEC and EHEC also disrupt Rho
GTPases to prevent internalization through the effector
EspH, which inactivates endogenous guanine nucleotide
exchange factors (GEFs), preventing Rho GTPase activa-
tion and thus inhibiting phagocytosis [46]. EspB, which
also functions in the translocation pore, interacts with
myosin family proteins inhibiting their interaction with
actin [47]. This disruption of the actomyosin system
results in inhibition of phagocytosis. EspF is central in
inhibiting internalization of non-opsonized bacteria and
in preventing transcytosis through M cells [33,48]. Al-
though EspF has been shown to inhibit PI 3 kinase
(PI3K)-dependent internalization through its N-terminal
101 residues, it does not inhibit opsonophagocytosis [49].
Several binding partners for EspF have been identified
including actin, profilin, sorting nexin 9 (SNX-9), and
Review Trends in Microbiology August 2013, Vol. 21, No. 8

Species inhibing internalizaon Species promong internalizaon

IgG
EPEC
EHEC

Yersinia Shigella Salmonella

FcγR EspF EspB

EspJ
Myosin Internalizaon
Internalizaon Internalizaon
YopH
Disrupon of
p85 P p85
Shigella endomembrane
YopO Fyb P Fyb trafficking
Cas P Cas IpaB IpaC
GDP EspH
YopT
Vacuolar lysis SifA

Salmonella
GDP GTP
Acn
polymerizaon Shigella Sifs

YopE

TRENDS in Microbiology

Figure 2. Disruption of phagocytosis and cell invasion. EPEC/EHEC (effectors in blue) and Yersinia (effectors in pink) inhibit internalization by disrupting phagocytic
signaling. In EPEC/EHEC, EspB prevents the interaction between actin and myosin, whereas EspH inhibits mammalian GEFs disrupting GTPase activation and actin
polymerization. EspF inhibits EPEC/EHEC internalization and EspJ inhibits opsonophagocytosis through unknown mechanisms. In Yersinia, YopH dephosphorylates host
proteins including p130Cas, p85, and Fyb to inhibit signal propagation. YopE, YopO, and YopT inhibit RhoGTPases by promoting GTP hydrolysis, sequestering the inactive
form and cleaving prenylyl groups, respectively. Shigella (in orange) and Salmonella (in green) trigger actin polymerization to promote their internalization. Once
internalized, Shigella disrupts the phagosome dependent on IpaB and IpaC to survive extracellularly. Salmonella prevents phagosome maturation through SifA-dependent
disruption of endomembrane signaling (see Figure 1 for further details). Abbreviations: EPEC/EHEC, enteropathogenic and enterohemorrhagic Escherichia coli; GEFs,
guanine nucleotide exchange factors; Fyb, Fyn binding protein; Sif, Salmonella induced filament.

neural Wiskott–Aldrich syndrome protein (N-WASP), but
the mechanism of action required to inhibit internaliza-
tion remains unclear [50,51]. Additionally, EspJ is impli-
cated in the inhibition of opsonophagocytosis pathways,
but it does not prevent internalization of uncoated bacte-
ria [48]. These pathogens therefore use an extensive array
of strategies to manipulate phagocytosis either by impair-
ing signaling to evade uptake or by facilitating their own
entry while blocking phagosomal maturation to establish
an intracellular niche.
Manipulation of cell survival
Programmed cell death through apoptosis is a tightly
regulated process activated by either intrinsic (intracellu-
lar: ER and mitochondria-mediated) or extrinsic (extracel-
lular: receptor-mediated) pathways. Both result in the
activation of the caspase cascade, which ultimately leads
to cell shrinkage, chromatin condensation, membrane
blebbing, and formation of vesicular buds known as apo-
ptotic bodies [52]. Pathogenic bacteria have developed the
ability to advantageously manipulate and avert host cell
death defense mechanisms to promote colonization and
infection by injecting specific T3SS effectors (Figure 3).
Inhibition of apoptosis
The EPEC/EHEC NleH family, homologs of Shigella OspG,
blocks apoptosis stimulated with staurosporine, brefeldin A,
and tunicamycin [53]. Treatment with brefeldin A and
tunicamycin induces the intrinsic apoptotic pathway lead-
ing to the conformational change and translocation of Bax to
the mitochondria, mitochondrial permeabilization, and cy-
tochrome C release. Through direct interaction with Bax
inhibitor I (BI-I), NleH blocks the intrinsic pathway [53].
Similarly, the Shigella effector IpgD blocks staurosporine-
induced apoptosis [54] by phosphorylating and stabilizing
the double minute 2 protein (MDM2). MDM2 represses p53
through binding its trans-activation domain [55]. Inhibition
of p53 blocks NF-kB–p53 proapoptotic signaling, inhibiting
apoptosis. Furthermore, the Shigella effector VirA promotes
further degradation of p53 via activation of calpain. Al-
though VirA has an antiapoptotic role, it also drives epithe-
lial cell necrosis and cell detachment in Shigella-infected
cells [55]. More recently, the EPEC/EHEC effector NleF has
been shown to specifically bind and inhibit the activity of
caspase-4, -8, and -9, inhibiting staurosporine- and TRAIL-
induced cell death, when transiently expressed [56].
Salmonella inhibits apoptosis and activates pro-surviv-
al signals, dependent on the effectors AvrA and SopB,
respectively. SopB sustains Akt activation and signaling
through the pro-survival PI3K–Akt pathway. Infection
with Salmonella enterica serovar Typhimurium (S. Typhi-
murium) sopB deletion mutant increases caspase-3 cleav-
age and apoptosis [57]. The antiapoptotic role of AvrA is
dependent on its acetyltransferase activity and inhibition
433
 Review Trends in Microbiology August 2013, Vol. 21, No. 8

EPEC
EHEC
Survival
signal
Salmonella Shigella Extrinsic
TRAIL, FasL
SopB NIeH
PI3K
Proinflammatory Caspase-8
signal BI-1
SipB
AKT

P
t-Bid
Bad Bax
MKK4/7
Caspase-2 NleF

AvrA
P Mitochondria
JNK EspF

IpgD Cyt C
NleF NleF
APAF-1
P EspH Caspase-9
MDM2
Caspase-4
EspM2 Caspase-3
p65/RelA SopE EspT Cif
Stress
p50 Abcf-2
Apoptosis
p53
EspF
Nucleus Calpain IpgD ER

TRENDS in Microbiology

Figure 3. Manipulation of cell survival. Intrinsic, extrinsic, and proinflammatory pathways involved in apoptosis are targeted by an extensive array of T3SS effectors. The EPEC/
EHEC (blue) NleH family blocks the intrinsic apoptotic pathway through direct interaction with BI-I. NleF binds caspase-4, -8, and -9, inhibiting both intrinsic and extrinsic
pathways. EspF induces cytochrome C (Cyt C) release from the mitochondria and degrades Abcf2 leading to the activation of caspase-9 and -3. Cif and EspH induce caspase-3
cleavage. Caspase-3 cleavage and cell death are recovered by the RhoGEF mimics EspM2, EspT, and SopE (from Salmonella, in green). Salmonella inhibits apoptosis through
SopB and AvrA. SopB sustains Akt activation and AvrA inhibits JNK proapoptotic signaling at the level of MKK4/7. In macrophages, SipB induces caspase-2 activation leading to
cell death. The Shigella effector IpgD blocks apoptosis by phosphorylating and stabilizing the MDM2. Furthermore, VirA promotes further degradation of p53 via activation of
calpain. Abbreviations: EPEC/EHEC, enteropathogenic and enterohemorrhagic Escherichia coli; T3SS, type III secretion system; BI-I, Bax inhibitor I; GEF, guanine nucleotide
exchange factor; JNK, c-Jun NH2-terminal kinase; MMK4/7, mitogen-activated protein kinase kinase 4/7; MDM2, double minute 2 protein.

of c-Jun NH2-terminal kinase (JNK) proapoptotic signal-
ing at the level of mitogen-activated protein kinase kinase
4 (MKK4) and MKK7 [58].
Induction of cell death
Although Salmonella, Shigella, Yersinia, EPEC, and
EHEC secrete T3SS effectors to block apoptosis, cell death
is apparent in in vivo models and in some in vitro studies.
EspF targets the mitochondria during EPEC infection of
HeLa cells inducing a loss of mitochondrial outer mem-
brane potential, cytochrome C release, and activation of
intrinsic apoptosis [59]. EspF binds Abcf2, a protein be-
longing to the ABC transporter family. During infection,
Abcf2 is degraded in an EspF-dependent manner leading to
an increase in caspase-9 and caspase-3 cleavage [59]. The
EPEC/EHEC effector Cif blocks cell cycle progression and
was recently identified as a late inducer (48 h) of EPEC-
induced apoptosis inducing caspase-3 cleavage and lactate
dehydrogenase (LDH) release [60]. The role of Cif in apo-
ptosis may be linked to its function in the inhibition of
the ubiquitin/proteasome pathway. EspH also induces
434
caspase-3 cleavage and cell death, which is recovered by
coexpression of the RhoGEF mimics EspM2, EspT (EPEC/
EHEC), and SopE (Salmonella) [61].
Furthermore, infection of macrophages with Salmonel-
la induces caspase-2 cleavage in a SipB-dependent man-
ner, leading to apoptosis and cell death independent of
caspase-1 [62].
Inflammatory response
Extracellular and intracellular pattern recognition recep-
tors (PRRs) recognize pathogen-associated molecular pat-
terns (PAMPs), which stimulate inflammatory signaling
cascades such as the NF-kB and MAPK pathways. Bacte-
rial pathogens have developed multiple mechanisms to
disrupt these pathways as outlined below.
Altering signaling to NF-kB
EPEC/EHEC, NleE, NleB, NleC, NleH, and Tir are immu-
nosuppressor T3SS effectors acting cooperatively to target
different proteins in the NF-kB signaling pathway
(Figure 4). NleE has methyltransferase activity and
 Review Trends in Microbiology August 2013, Vol. 21, No. 8

EPEC
EHEC TNF-α IL-1β

Shigella
Salmonella Yersinia TNF-α R IL-1β R

Ospl Ubc13
TRAF2 TRAF6
SipA GAPDH ub Tir SH1 ub
EspT IpGB2 NOD1/2 NleB
SopE SopE2 NleE
SopB
TAB2/3
ABIN-1
AvrA
GDP GTP IpaH9.8
SspH1 SseL TAK1

NEMO YopP/J
IKKγ
P
E2–E3 OspG
P complex P Proteosomal
IκBα IκBα degradaon
p65/RelA ub
p50 NleC
RPS3
OspZ
YopE

NleH1
p65/RelA
RPS3
p50
Nucleus
TRENDS in Microbiology

Figure 4. NF-kB subversion. NleB, NleE, NleH1, Tir, and NleC from EPEC/EHEC (in blue) inhibit NF-kB activation by targeting TRAF2, TAB1/2, RPS3, TRAF6, or p65 directly,
respectively. NleB reduces TRAF2 polyubiquitination by blocking its interaction with GAPDH. NleE blocks TAK2/3 activation. NleH1 binds and blocks the translocation of
RPS3 into the nucleus. Through its ITIM-like sequence that recruits SHP-1 to bind TRAF6, Tir blocks TRAF6 activation. NleC cleaves p65. Shigella affects NF-kB signaling
through OspG, OspI, IpaH9.8, and OspZ (in orange). OspG interferes with the degradation of IkB-a, OspI blocks the polyubiquitination of TRAF6, IpaH9.8 promotes the
degradation of NEMO, and OspZ blocks the translocation of p65 through an unknown mechanism. Yersinia translocates YopE (in pink), which inhibits activation of NF-kB,
and YopP/J that acts as a serine/threonine acetyltransferase targeting TAK1. SseL, SspH1, and AvrA (in green) secreted by Salmonella have been identified to inhibit NF-kB
signaling pathways. However, the RhoGEF mimics SopE, SopE2, EspT, and IpgB2, as well as the PI phosphatase SopB, induce NF-kB activation through their impact on
RhoGTPases. In addition, SipA activates NF-kB through NOD1. Abbreviations: NF-kB, nuclear factor-kB; EPEC/EHEC, enteropathogenic and enterohemorrhagic Escherichia
coli; T3SS, type III secretion system; RPS3, ribosomal protein S3; TRAF, TNF receptor-associated factor; TAB, TAK1 binding protein; TAK, TGFb activated kinase; ITIM,
immunoreceptor tyrosine-based inhibition motif; IkB-a, inhibitor of kB, NEMO, NF-kB essential modulator;GAPDH, glyceraldehyde 3-phosphate dehydrogenase; GEF,
guanine nucleotide exchange factor; PI, phosphoinositide.

specifically methylates TAB2/3 to block TRAF6-induced
activation of the TAK1–NF-kB pathway in response to
tumor necrosis factor-a (TNF-a) and interleukin-1b (IL-
1b) [63–65]. NleB has N-acetyl-D-glucosamine transferase
activity and modifies glyceraldehyde 3-phosphate dehydro-
genase (GAPDH) to disrupt the TRAF2–GAPDH interac-
tion and reduce TRAF2 polyubiquitination, which is
needed for NF-kB activation [66]. Through its consensus
zinc metalloprotease motif, NleC has been shown to cleave
the NF-kB p65 subunit (RelA), blocking the production of
IL-8 following infection [67]. Although both NleH1 and
NleH2 bind to the ribosomal protein S3 (RPS3) subunit
of NF-kB transcriptional complexes, NleH1, but not
NleH2, blocks translocation of NF-kB into the nucleus,
without affecting IkB-a degradation [68,69]. The effector
Tir, most well characterized for its role in pedestal forma-
tion, is also able to inhibit NF-kB activation in response to
TNF-a stimulation. Tir has an immunoreceptor tyrosine-
based inhibition motif (ITIM)-like sequence that recruits
SHP-1 to bind TRAF6 and inhibits cytokine production in
cultured cells and in vivo [70,71].
Although Shigella peptidoglycan stimulates signaling
pathways required for an efficient inflammatory response,
which is thought to be important for the infection strategy
of Shigella, studies have also identified Shigella T3SS
effectors which affect the NF-kB pathway, including OspZ
and OspG, homologs of NleE and NleH, respectively. Simi-
lar to NleE, OspZ blocks the translocation of NF-kB to the
nucleus [64], but whether OspZ shares the same cysteine
methylase activity of NleE is still unknown. Through its
impact on the E2–E3 ubiquitination complex, OspG inter-
feres with the ubiquitin-dependent proteasomal degrada-
tion of IkB-a [72,73], in contrast to NleH. OspG also
displays kinase activity, inducing its autophosphorylation
435
Review
which is necessary for its function. The IpaH effectors are
found multiple times in Shigella spp. and the crystal
structure suggests that they belong to a new class of E3
ubiquitin ligases, with a role in suppressing innate im-
mune responses [74]. IpaH9.8 dampens NF-kB signaling
by interacting with NEMO/IKK and ABIN-1. ABIN-1 pro-
motes polyubiquitination of NEMO, which then undergoes
proteasome-dependent degradation, ultimately blocking
NF-kB activation [75]. OspI selectively deamidates a glu-
tamine residue in the E2 enzyme UBC13 to abolish its
ubiquitin-conjugating activity, which is required for acti-
vation of the TRAF6–NF-kB pathway [76].
Similarly, YopE, YopP, and YopJ (in Yersinia enteroco-
litica, Yersinia pestis, and Yersinia pseudotuberculosis re-
spectively) target the NF-kB pathway. YopE inhibits
activation of NF-kB, which in part prevents the production
of IL-8 [77]. YopJ acts as a cysteine protease and was
reported to hydrolyze ubiquitin and SUMO [78]. YopP/J
also functions as a serine/threonine acetyltransferase tar-
geting the activation loop of Drosophila TAK1, which
blocks its phosphorylation and subsequent kinase activa-
tion [79].
Despite the profound proinflammatory nature of Sal-
monella, the effectors SseL [80], SspH1 [81], and possibly
AvrA [82] have been found to inhibit NF-kB signaling
pathways.
Targeting MAPK
MAPK, with the extracellular signal-regulated protein
kinases (ERKs), the p38 MAP kinases, and the JNKs
cooperate with other signaling pathways to regulate the
expression of inflammatory genes [83]. In addition to NF-
kB, MAPK is also targeted by T3SS effectors to dampen the
inflammatory response. The EPEC effector NleD is a zinc-
dependent metalloprotease specifically targeting MAP
kinases JNK and p38 but not ERK, thereby blocking
nuclear translocation of the AP-1 transcription factor,
which regulates multiple cell processes including inflam-
mation [84]. NleC affects the p38 pathway and plays a
significant role in the virulence of C. rodentium in vivo [85].
Moreover, deletion of Tir enhanced the activation of ERK,
JNK, and p38 3 h post-EPEC infection [71].
Although it is not clear how nuclear IpaH9.8 from
Shigella flexneri affects MAPK signaling, its ubiquitin
ligase activity targets the yeast MAPK Ste7 to interrupt
pheromone response signaling by promoting the protea-
some-dependent destruction of Ste7 [86]. OspF has phos-
phothreonine lyase activity and translocates to the
nucleus, mediating irreversible dephosphorylation and
hence inactivation of MAPKs [87]. This also inhibits down-
stream phosphorylation of histone H3 at Ser10, altering
chromatin accessibility of the IL-8 promoter to NF-kB in
epithelial cells [88].
Infection of HeLa cells with the Yersinia DyopETHJ
mutant expressing either YopE or YopT indicates that
YopE can strongly inhibit activation of JNK, ERK, and
prevent production of IL-8, whereas YopT moderately
inhibited these responses [77]. YopH inactivates the
PI3K–AKT cascade, which in macrophages correlated with
the downregulation of mRNA coding for monocyte che-
moattractant protein 1 (MCP-1) [89]. YopP/J also represses
436
Trends in Microbiology August 2013, Vol. 21, No. 8
the MAPK signaling pathway by an unknown mechanism
[90].
Salmonella AvrA blocks JNK activation at the level of
the MAPK kinases MKK4 and MKK7 in both AvrA trans-
genic Drosophila and murin cells. Consistent with this
finding, a DavrA mutant induces higher levels of JNK
activation and inflammation in infected murine intestine
[58]. Interestingly, the ERK pathway, which is activated
upon S. Typhimurium infection, is required for the phos-
phorylation of AvrA, which then impairs the JNK pathway
through its interaction with MKK7, but not the p38 or NF-
kB pathways. In addition, SpvC, a member of the OspF
family, has been shown to abrogate the activation of the
ERK, JNK, and p38 kinases [87].
Upregulation of inflammatory responses
Bacterial RhoGEF mimics [1] have a well-defined role in
cytoskeleton remodeling and also induce innate immune
responses. S. Typhimurium induces inflammation in cul-
tured epithelial cells through SopE and SopE2 [91]. SopE
and SopE2 are GEFs for Rac1, Cdc42, RhoA, and RhoG [1].
SopB is a PI phosphatase, which activates RhoG and Cdc42
by stimulating their endogenous exchange factors [25], and
also induces inflammation [91]. Activation of these
GTPases by SopE, SopE2, and SopB leads to stimulation
of NF-kB, ERK, JNK, and p38 pathways independently of
the intracellular sensors NOD1, NOD2, and RIP2 kinase
[91]. However, it has recently been shown that SopE
triggers the NOD1–RIP2–NF-kB pathway [92]. It has been
suggested that the induction of inflammation is crucial for
the ability of Salmonella to grow in the intestinal tract,
because essential nutrients do not become available at this
site unless such responses are stimulated [93]. Moreover,
recent reports indicated that Salmonella is able to use
electron acceptors generated during inflammation in the
gut, thereby gaining a competitive advantage over com-
mensal bacteria [94,95]. The EPEC RhoGEF mimic EspT
induces production of IL-8 and IL-1b in macrophages
through Rac1 [96], whereas the Shigella RhoGEF mimic
IpgB2 activates NF-kB via a GEF-H1, RhoA, and NOD1
pathway [97], further strengthening the link between the
small Rho GTPases/RhoGEF mimic superfamily and im-
mune responses. Salmonella SipA, a non-RhoGEF mimic,
enhances mucosal inflammation in the mouse colitis model
and NF-kB activation in cultured cells in a NOD1/NOD2-
dependent manner [98]. Given the central role MAPKs and
NF-kB play in antimicrobial signaling, it is not surprising
that bacterial pathogens have evolved many mechanisms
to affect these pathways.
Targeting inflammasomes
The inflammasome is a key component of cytosolic surveil-
lance in macrophages where the activated NOD-like recep-
tor (NLR) protein family (e.g., NLRC4 and NLRP3) recruits
the adaptor protein apoptosis-associated speck-like pro-
tein containing a caspase recruitment domain (ASC),
which in turn recruits and activates caspase-1. The acti-
vated caspase-1 regulates maturation of the proinflamma-
tory cytokines IL-1b and IL-18 and the rapid inflammatory
form of cell death, called pyroptosis, characterized by the
formation of pores in the plasma membrane and activation
Review Trends in Microbiology August 2013, Vol. 21, No. 8

Table 1. Summary of the activity of the T3SS effectors discussed

T3SS Putative or Putative or known Homolog Associated function Refs
effector known host enzymatic activity
partner
Attaching and NleE TAB2, TAB3 Methyltransferase OspZ Inhibits NF-kB activation [63–65]
effacing (A/E) and inflammatory
pathogens response
NleB GAPDH Glycosyltransferase SseK Inhibits TRAF2–NF-kB [64,66]
activation
EspI Sec24 – Disrupts secretion by [3,4,6]
(NleA) reducing ER–Golgi
trafficking; alters tight
junctions
NleC RelA Zinc-metalloprotease Cleaves RelA; affects the [84,85]
p38 pathway
NleD JNK and p38 Zinc-metalloprotease Degrades JNK and p38 [84]
and blocks the
translocation of AP-1
EspG Rabs, ARFs, PAKs, Rab-GAP VirA Disrupts secretion; [8–11]
GM130, RACK1 inhibits IL-8 secretion
NleH1/ Bax inhibitor I, Serine threonine OspG Blocks the intrinsic [53,68,69]
NleH2 RPS3 kinase apoptotic pathway; NleH1
blocks the translocation of
RPS3 into the nucleus
EspF Abcf2, actin, – TccP/EspFu Disrupts mitochondria; [33,49–51,59]
profilin, SNX-9, inhibits non-opsonized
N-WASP bacteria internalization
and transcytosis through
M cells; inhibition of PI3K;
increases caspase-9 and -3
activation
Tir SHP-1 Has an ITIM-like Blocks TRAF6–NF-kB [70,71]
sequence pathway and inhibits
cytokine production;
blocks MAPK signaling
Cif – – Induces caspase-3 [60]
cleavage; inhibits the
ubiquitin/proteasome
pathway
EspH DH–PH – Inhibits host RhoGEFs; [46]
domain induces caspase-3
RhoGEFs activation; inhibits
phagocytosis
EspB Myosin – Inhibits myosin/actin [47]
interaction; blocks
phagocytosis
EspJ – – Inhibits [48]
opsonophagocytosis
EspT Rac1, cdc42 RhoGEF mimics RhoGEF Invasion; induces IL-8 and [32,96]
mimics IL-1b production in
macrophages
NleF Tmp21, – May affect ER–Golgi and [7,56]
caspase-4/-5 Golgi–PM trafficking;
and -9 inhibits caspase-4/-5/-9
Shigella VirA Rabs, ARFs Rab-GAP EspG Activates calpain; reduces [55]
autophagic recognition
IpgD PI(4,5)P2, PI phosphatase Activates Akt; stabilizes [54]
MDM2 MDM2 and inhibits p53
proapoptotic signaling
IpaC – – Lyses the Shigella vacuole [21]
IpaB Caspase-1 – SipB Activates caspase-1 and [20,21,105]
pyroptosis; causes
Shigella entry into
epithelial cells and escape
from the phagocytic
vacuole
OspZ – Methyltransferase NleE Blocks NF-kB [64]
translocation
OspG Binds various Kinase NleH Blocks the E2–E3 [72,73]
ubiquitin- ubiquitination complex

437
Review Trends in Microbiology August 2013, Vol. 21, No. 8

Table 1 (Continued )
T3SS Putative or Putative or known Homolog Associated function Refs
effector known host enzymatic activity
partner
conjugating enzymes and alters IkB-a
belonging to the degradation
SCFb–TrCP complex
IpaH9.8 NEMO/IKK, ABIN-1 E3 ubiquitin ligases Dampens NF-kB signaling; [74,75,86]
blocks the yeast MAPK
Ste7
OspI UBC13 Glutamine deamidase Deamidates UBC13 and [76]
blocks the TRAF6–NF-kB
pathway
OspF MAPK Phospho-threonine SpvC Dephosphorylates MAPKs [87,88]
lyase and inhibits the
downstream
phosphorylation of
histone H3 at Ser10,
altering the chromatin
accessibility of the IL-8
promoter to NF-kB
IpgB2 RhoA RhoGEF mimics RhoGEF Activates NF-kB [97]
mimics
IpaJ ARF1 Cysteine protease Cleaves the myristoyl [12]
group from ARF1 and
alters the host secretory
pathway
Yersinia spp. YopJ/P IKKb Serine/threonine AvrA Inhibits NF-kB/MAPK pro- [78,79,
acetyltransferase, survival signaling; induces 106,107]
cysteine protease apoptosis, pyroptosis and
IL-1b secretion; hydrolyzes
ubiquitin and SUMO
YopK/ – – Inhibits ASC-dependent [108]
YopQ inflammasome activation
YopH p130Cas, Fyb, FAK, Tyrosine phosphatase Dephosphorylates p130C [35–39,89]
p85, Lyn, paxillin as required for Yersinia
internalization; inhibits
PI3K–AKT in
macrophages, and MCP-1
expression
YopE Rac1 RhoGAP Inhibits NF-kB activation [77,109]
and IL-8 production;
inactivates Rac1; inhibits
JNK and ERK activation;
inhibits caspase-1
activation
YopO/ Rac1 GDI-like domain Inactivates Rac1 and [43,44]
YpkA blocks phagocytosis
YopM Caspase-1 Inhibits inflammasome [110]
assembly
YopT RhoA, Rac1, Cysteine protease Inactivates RhoA, Rac1, [45,77]
and Cdc42 and Cdc42; moderately
inhibits JNK and ERK
Salmonella spp. SipC Exo70 – Provides extra membrane [13]
to facilitate invasion
SifA SKIP, RhoA-GDP – Excludes lysosomal [16]
hydrolases from SCV
SseF – – Produces Sifs [18,19]
SseG – – Produces Sifs [18,19]
SopB PI(4,5)P2 PI phosphatase Facilitates invasion and [25,57,91]
(SigD) prevents phagolysosome
formation; sustains PI3K–
AKT antiapoptotic
signaling; activates RhoG
and Cdc42; induces NF-kB
and MAPKs
AvrA MKK4, MKK7, IkB-a Acetyltransferase; YopJ/P Inhibits JNK proapoptotic [58,82]
deubiquitinase signaling; inhibits NF-kB
activation and host
immune response
SipB Caspase-1 – IpaB [62,104]

438
Review
Table 1 (Continued )
T3SS Putative or Putative or known
effector known host enzymatic activity
partner
SseL IkB-a Deubiquitinase
SspH1 PKN1 – Blocks NF-kB activation
SpvC MAPK Phospho-threonine lyase
SopE/SopE2 Rho GTPases RhoGEF
SipA – –
PrgJ – –
of an unidentified endonuclease [99]. Because IL-1b tran-
scription is highly regulated by NF-kB, all the described
effectors affecting the NF-kB pathway could affect IL-1b
expression and hence the pool of IL-1b available for cas-
pase-1 cleavage.
S. Typhimurium activates caspase-1 in macrophages in a
SPI-1 T3SS-dependent manner [100]. In addition to flagellin
and SopE [101], which induce caspase-1 activation, PrgJ,
which forms the periplasmic rod of the T3SS needle complex,
is recognized by NLRC4 [102]. During systemic infection,
Salmonella spp. downregulate expression of the flagellin,
SopE, and PrgJ, while simultaneously upregulating the
SPI-2 T3SS, which is not detected by NLRC4. The Salmo-
nella T3SS also appears to inhibit pyroptosis in B cells
through downregulation of the NLRC4 inflammasome
and inhibition of caspase-1 cleavage, although the effector
responsible has not been identified [103].
The Shigella T3SS effector IpaB and its Salmonella
homolog SipB induce cell death in macrophages with fea-
tures of pyroptosis by binding and activating caspase-1
[104,105]. Similarly, although YopP/J was originally de-
scribed as inducing apoptosis in macrophages through the
MAPK and NF-kB pathways [106], a recent paper demon-
strated that it induces pyroptosis and IL-1b secretion
during infection through caspase-1 [107]. In contrast to
this, YopK/YopQ of Yersinia block pyroptosis by preventing
ASC-dependent inflammasome activation in response to
the T3SS, promoting bacterial survival and replication.
Expression of YopK is insufficient to inhibit YopJ-induced
pyroptosis suggesting that they act on distinct pathways of
caspase-1 activation [108]. YopE and YopT of Y. enteroco-
litica negatively regulate caspase-1 by targeting Rac1
[109], and YopM blocks the assembly of the inflammasome
by directly targeting caspase-1 [110]. Induction of the
inflammasome and pyroptosis in macrophages may be a
mechanism by which pathogens preferentially trigger im-
mune cell death, resulting in bacterial dissemination and
disruption of host innate immune signaling.
Concluding remarks
Recent advances have enhanced our understanding of how
T3SS effectors manipulate host cell processes enabling
Trends in Microbiology August 2013, Vol. 21, No. 8
Homolog Associated function Refs
Activates caspase-1 and
pyroptosis; activation of
caspase-2-dependent
apoptosis
Impairs IkB-a degradation [80]
and NF-kB activation
[81]
OspF Blocks MAPK signaling [87]
RhoGEF mimics Activates NF-kB, ERK, [1,91,92,101]
JNK, and p38 pathways;
induces caspase-1
activation
Induces NF-kB activation [98]
in a NOD1/NOD2-
dependent manner
Is recognized by NLRC4 [102]
bacterial colonization, multiplication, and evasion of in-
nate immunity and host responses. Here, we have
reviewed the mechanisms used by EPEC/EHEC, Yersinia,
Salmonella, and Shigella to disrupt key cellular signaling
pathways of early host defense processes (Table 1). It is
evident that although they have evolved diverse lifestyles
within the host (i.e., intracellular and extracellular), these
pathogens share common strategies to counteract host
defenses and use specific effectors to mitigate cell-specific
obstacles. For example, whereas inhibition of apoptosis in
epithelial cells is beneficial to promote colonization, induc-
tion of pyroptosis in macrophages may facilitate dissemi-
nation. Another common theme is the existence of a high
level of synergy between different T3SS effectors that
together inhibit or activate specific signaling pathways,
while at the same time other effectors antagonize the
activity of each other. Accordingly, great care should be
taken when dissecting the function of individual effectors
and their contribution to subversion of host cell signaling.
Acknowledgments
We are grateful for the financial support from the Biotechnology and
Biological Sciences Research Council (BBSRC) and the Medical Research
council (MRC).
References
1 Bulgin, R. et al. (2010) Bacterial guanine nucleotide exchange factors
SopE-like and WxxxE effectors. Infect. Immun. 78, 1417–1425
2 Faini, M. et al. (2013) Vesicle coats: structure, function, and general
principles of assembly. Trends Cell Biol. 23, 279–288
3 Kim, J. et al. (2007) Biogenesis of g-secretase early in the secretory
pathway. J. Cell Biol. 179, 951–963
4 Thanabalasuriar, A. et al. (2012) Sec24 interaction is essential for
localization and virulence-associated function of the bacterial effector
protein NleA. Cell. Microbiol. 14, 1206–1218
5 Mundy, R. et al. (2004) Distribution of espI among clinical
enterohaemorrhagic and enteropathogenic Escherichia coli isolates.
J. Med. Microbiol. 53, 1145–1149
6 Thanabalasuriar, A. et al. (2010) The bacterial virulence factor NleA is
required for the disruption of intestinal tight junctions by
enteropathogenic Escherichia coli. Cell. Microbiol. 12, 31–41
7 Olsen, R.L. et al. (2013) The enterohemorrhagic Escherichia coli effector
protein NleF binds mammalian Tmp21. Vet. Microbiol. 64, 164–170
8 Selyunin, A.S. et al. (2011) The assembly of a GTPase-kinase
signalling complex by a bacterial catalytic scaffold. Nature 469,
107–111
439
Review
9 Clements, A. et al. (2011) EspG of enteropathogenic and
enterohemorrhagic E. coli binds the Golgi matrix protein GM130 and
disrupts the Golgi structure and function. Cell. Microbiol. 13, 1429–1439
10 Dong, N. et al. (2012) Structurally distinct bacterial TBC-like GAPs
link Arf GTPase to Rab1 inactivation to counteract host defenses. Cell
150, 1029–1041
11 Germane, K.L. and Spiller, B.W. (2011) Structural and functional
studies indicate that the EPEC effector, EspG, directly binds p21-
activated kinase. Biochemistry 50, 917–919
12 Burnaevskiy, N. et al. (2013) Proteolytic elimination of N-
myristoylmodifications by the Shigella virulence factor IpaJ.
Nature 496, 106–109
13 Nichols, C.D. and Casanova, J.E. (2010) Salmonella-directed
recruitment of new membrane to invasion foci via the host exocyst
complex. Curr. Biol. 20, 1316–1320
14 Meresse, S. et al. (1999) Controlling the maturation of pathogen-
containing vacuoles: a matter of life and death. Nat. Cell Biol. 1,
E183–E188
15 Meresse, S. et al. (1999) The rab7 GTPase controls the maturation of
Salmonella typhimurium-containing vacuoles in HeLa cells. EMBO J.
18, 4394–4403
16 McGourty, K. et al. (2012) Salmonella inhibits retrograde trafficking
of mannose-6-phosphate receptors and lysosome function. Science
338, 963–967
17 Drecktrah, D. et al. (2007) Salmonella trafficking is defined by
continuous dynamic interactions with the endolysosomal system.
Traffic 8, 212–225
18 Drecktrah, D. et al. (2008) Dynamic behavior of Salmonella-induced
membrane tubules in epithelial cells. Traffic 9, 2117–2129
19 Rajashekar, R. et al. (2008) Dynamic remodeling of the endosomal
system during formation of Salmonella-induced filaments by
intracellular Salmonella enterica. Traffic 9, 2100–2116
20 Blocker, A. et al. (1999) The tripartite type III secreton of Shigella
flexneri inserts IpaB and IpaC into host membranes. J. Cell Biol. 147,
683–693
21 High, N. et al. (1992) IpaB of Shigella flexneri causes entry into
epithelial cells and escape from the phagocytic vacuole. EMBO J.
11, 1991–1999
22 Fernandez-Prada, C.M. et al. (2000) Shigella flexneri IpaH(7.8)
facilitates escape of virulent bacteria from the endocytic vacuoles of
mouse and human macrophages. Infect. Immun. 68, 3608–3619
23 Mounier, J. et al. (1999) Rho family GTPases control entry of Shigella
flexneri into epithelial cells but not intracellular motility. J. Cell Sci.
112, 2069–2080
24 Zhou, D. and Galan, J. (2001) Salmonella entry into host cells: the
work in concert of type III secreted effector proteins. Microbes Infect.
3, 1293–1298
25 Marcus, S.L. et al. (2002) Salmonella enterica serovar Typhimurium
effector SigD/SopB is membrane-associated and ubiquitinated inside
host cells. Cell. Microbiol. 4, 435–446
26 Ramel, D. et al. (2011) Shigella flexneri infection generates the lipid
PI5P to alter endocytosis and prevent termination of EGFR signaling.
Sci. Signal. 4, ra61
27 Niebuhr, K. et al. (2002) Conversion of PtdIns(4,5)P(2) into PtdIns(5)P
by the S. flexneri effector IpgD reorganizes host cell morphology.
EMBO J. 21, 5069–5078
28 Peiser, L. et al. (2000) Macrophage class A scavenger receptor-
mediated phagocytosis of Escherichia coli: role of cell
heterogeneity, microbial strain, and culture conditions in vitro.
Infect. Immun. 68, 1953–1963
29 Anderson, C.L. et al. (1990) Phagocytosis mediated by three distinct Fcg
receptor classes on human leukocytes. J. Exp. Med. 171, 1333–1345
30 Ross, G.D. et al. (1992) Macrophage cytoskeleton association with CR3
and CR4 regulates receptor mobility and phagocytosis of iC3b-
opsonized erythrocytes. J. Leukoc. Biol. 51, 109–117
31 Isberg, R.R. (1990) Pathways for the penetration of enteroinvasive
Yersinia into mammalian cells. Mol. Biol. Med. 7, 73–82
32 Bulgin, R. et al. (2009) The T3SS effector EspT defines a new category
of invasive enteropathogenic E. coli (EPEC) which form intracellular
actin pedestals. PLoS Pathog. 5, e1000683
33 Martinez-Argudo, I. et al. (2007) Translocation of enteropathogenic
Escherichia coli across an in vitro M cell model is regulated by its type
III secretion system. Cell. Microbiol. 9, 1538–1546
440
Trends in Microbiology August 2013, Vol. 21, No. 8
34 Goosney, D.L. et al. (1999) Enteropathogenic Escherichia coli inhibits
phagocytosis. Infect. Immun. 67, 490–495
35 Black, D.S. et al. (1998) Identification of an amino-terminal substrate-
binding domain in the Yersinia tyrosine phosphatase that is required
for efficient recognition of focal adhesion targets. Mol. Microbiol. 29,
1263–1274
36 de la Puerta, M.L. et al. (2009) Characterization of new substrates
targeted by Yersinia tyrosine phosphatase YopH. PLoS ONE 4, e4431
37 Hamid, N. et al. (1999) YopH dephosphorylates Cas and Fyn-binding
protein in macrophages. Microb. Pathog. 27, 231–242
38 Persson, C. et al. (1997) The PTPase YopH inhibits uptake of Yersinia,
tyrosine phosphorylation of p130Cas and FAK, and the associated
accumulation of these proteins in peripheral focal adhesions. EMBO
J. 16, 2307–2318
39 Weidow, C.L. et al. (2000) CAS/Crk signalling mediates uptake of
Yersinia into human epithelial cells. Cell. Microbiol. 2, 549–560
40 Von Pawel-Rammingen, U. et al. (2000) GAP activity of the Yersinia
YopE cytotoxin specifically targets the Rho pathway: a mechanism
for disruption of actin microfilament structure. Mol. Microbiol. 36,
737–748
41 Andor, A. et al. (2001) YopE of Yersinia, a GAP for Rho GTPases,
selectively modulates Rac-dependent actin structures in endothelial
cells. Cell. Microbiol. 3, 301–310
42 Roppenser, B. et al. (2009) Yersinia enterocolitica differentially
modulates RhoG activity in host cells. J. Cell Sci. 122, 696–705
43 Prehna, G. et al. (2006) Yersinia virulence depends on mimicry of host
Rho-family nucleotide dissociation inhibitors. Cell 126, 869–880
44 Groves, E. et al. (2010) Sequestering of Rac by the Yersinia effector
YopO blocks Fcg receptor-mediated phagocytosis. J. Biol. Chem. 285,
4087–4098
45 Shao, F. et al. (2003) Biochemical characterization of the Yersinia
YopT protease: cleavage site and recognition elements in Rho
GTPases. Proc. Natl. Acad. Sci. U.S.A. 100, 904–909
46 Dong, N. et al. (2010) A bacterial effector targets host DH–PH domain
RhoGEFs and antagonizes macrophage phagocytosis. EMBO J. 29,
1363–1376
47 Iizumi, Y. et al. (2007) The enteropathogenic E. coli effector EspB
facilitates microvillus effacing and antiphagocytosis by inhibiting
myosin function. Cell Host Microbe 2, 383–392
48 Marches, O. et al. (2008) EspJ of enteropathogenic and
enterohaemorrhagic Escherichia coli inhibits opsono-phagocytosis.
Cell. Microbiol. 10, 1104–1115
49 Quitard, S. et al. (2006) The enteropathogenic Escherichia coli EspF
effector molecule inhibits PI-3 kinase-mediated uptake independently
of mitochondrial targeting. Cell. Microbiol. 8, 972–981
50 Peralta-Ramirez, J. et al. (2008) EspF interacts with nucleation-
promoting factors to recruit junctional proteins into pedestals for
pedestal maturation and disruption of paracellular permeability.
Infect. Immun. 76, 3854–3868
51 Alto, N.M. et al. (2007) The type III effector EspF coordinates
membrane trafficking by the spatiotemporal activation of two
eukaryotic signaling pathways. J. Cell Biol. 178, 1265–1278
52 Taylor, R.C. et al. (2008) Apoptosis: controlled demolition at the
cellular level. Nat. Rev. Mol. Cell Biol. 9, 231–241
53 Hemrajani, C. et al. (2010) NleH effectors interact with Bax inhibitor-
1 to block apoptosis during enteropathogenic Escherichia coli
infection. Proc. Natl. Acad. Sci. U.S.A. 107, 3129–3134
54 Clark, C.S. and Maurelli, A.T. (2007) Shigella flexneri inhibits
staurosporine-induced apoptosis in epithelial cells. Infect. Immun.
75, 2531–2539
55 Bergounioux, J. et al. (2012) Calpain activation by the Shigella
flexneri effector VirA regulates key steps in the formation and life
of the bacterium’s epithelial niche. Cell Host Microbe 11, 240–252
56 Blasche, S. et al. (2013) The E. coli effector protein NleF is a caspase
inhibitor. PLoS ONE 8, e58937
57 Knodler, L.A. et al. (2005) The Salmonella effector protein SopB
protects epithelial cells from apoptosis by sustained activation of
Akt. J. Biol. Chem. 280, 9058–9064
58 Jones, R.M. et al. (2008) Salmonella AvrA coordinates suppression of
host immune and apoptotic defenses via JNK pathway blockade. Cell
Host Microbe 3, 233–244
59 Nougayrede, J.P. and Donnenberg, M.S. (2004) Enteropathogenic
Escherichia coli EspF is targeted to mitochondria and is required to
Review
initiate the mitochondrial death pathway. Cell. Microbiol. 6,
1097–1111
60 Samba-Louaka, A. et al. (2009) The enteropathogenic Escherichia coli
effector Cif induces delayed apoptosis in epithelial cells. Infect.
Immun. 77, 5471–5477
61 Wong, A.R. et al. (2012) The interplay between the Escherichia coli
Rho guanine nucleotide exchange factor effectors and the mammalian
RhoGEF inhibitor EspH. mBio 3, e00250–11, (http://dx.doi.org/
10.1128/mBio.00250-11)
62 Jesenberger, V. et al. (2000) Salmonella-induced caspase-2 activation
in macrophages: a novel mechanism in pathogen-mediated apoptosis.
J. Exp. Med. 192, 1035–1046
63 Nadler, C. et al. (2010) The type III secretion effector NleE inhibits
NF-kB activation. PLoS Pathog. 6, e1000743
64 Newton, H.J. et al. (2010) The type III effectors NleE and NleB from
enteropathogenic E. coli and OspZ from Shigella block nuclear
translocation of NF-kB p65. PLoS Pathog. 6, e1000898
65 Vossenkamper, A. et al. (2010) Inhibition of NF-kB signaling in
human dendritic cells by the enteropathogenic Escherichia coli
effector protein NleE. J. Immunol. 185, 4118–4127
66 Gao, X. et al. (2013) NleB, a bacterial effector with glycosyltransferase
activity, targets GAPDH function to inhibit NF-kB activation. Cell
Host Microbe 13, 87–99
67 Yen, H. et al. (2010) NleC, a type III secretion protease, compromises
NF-kB activation by targeting p65/RelA. PLoS Pathog. 6, e1001231
68 Pham, T.H. et al. (2012) Functional differences and interactions
between the Escherichia coli type III secretion system effectors
NleH1 and NleH2. Infect. Immun. 80, 2133–2140
69 Gao, X. et al. (2009) Bacterial effector binding to ribosomal protein s3
subverts NF-kB function. PLoS Pathog. 5, e1000708
70 Ruchaud-Sparagano, M.H. et al. (2011) The enteropathogenic E. coli
(EPEC) Tir effector inhibits NF-kB activity by targeting TNFa
receptor-associated factors. PLoS Pathog. 7, e1002414
71 Yan, D. et al. (2012) Inhibition of TLR signaling by a bacterial protein
containing immunoreceptor tyrosine-based inhibitory motifs. Nat.
Immunol. 13, 1063–1071
72 Kim, D.W. et al. (2005) The Shigella flexneri effector OspG interferes
with innate immune responses by targeting ubiquitin-conjugating
enzymes. Proc. Natl. Acad. Sci. U.S.A. 102, 14046–14051
73 Zhou, Y. et al. (2013) The Shigella type three secretion system effector
OspG directly and specifically binds to host ubiquitin for activation.
PLoS ONE 8, e57558
74 Singer, A.U. et al. (2008) Structure of the Shigella T3SS effector IpaH
defines a new class of E3 ubiquitin ligases. Nat. Struct. Mol. Biol. 15,
1293–1301
75 Ashida, H. et al. (2010) A bacterial E3 ubiquitin ligase IpaH9.8 targets
NEMO/IKKg to dampen the host NF-kB-mediated inflammatory
response. Nat. Cell Biol. 12, 66–73
76 Sanada, T. et al. (2012) The Shigella flexneri effector OspI deamidates
UBC13 to dampen the inflammatory response. Nature 483, 623–626
77 Viboud, G.I. et al. (2006) Comparison of YopE and YopT activities in
counteracting host signalling responses to Yersinia
pseudotuberculosis infection. Cell. Microbiol. 8, 1504–1515
78 Orth, K. et al. (2000) Disruption of signaling by Yersinia effector YopJ,
a ubiquitin-like protein protease. Science 290, 1594–1597
79 Paquette, N. et al. (2012) Serine/threonine acetylation of TGFb-
activated kinase (TAK1) by Yersinia pestis YopJ inhibits innate
immune signaling. Proc. Natl. Acad. Sci. U.S.A. 109, 12710–12715
80 Le Negrate, G. et al. (2008) Salmonella secreted factor L
deubiquitinase of Salmonella typhimurium inhibits NF-kB,
suppresses IkBa ubiquitination and modulates innate immune
responses. J. Immunol. 180, 5045–5056
81 Haraga, A. and Miller, S.I. (2003) A Salmonella enterica serovar
typhimurium translocated leucine-rich repeat effector protein
inhibits NF-kB-dependent gene expression. Infect. Immun. 71,
4052–4058
82 Collier-Hyams, L.S. et al. (2002) Cutting edge: Salmonella AvrA
effector inhibits the key proinflammatory, anti-apoptotic NF-kB
pathway. J. Immunol. 169, 2846–2850
83 Zhang, Y.L. and Dong, C. (2005) MAP kinases in immune responses.
Cell. Mol. Immunol. 2, 20–27
84 Baruch, K. et al. (2011) Metalloprotease type III effectors that
specifically cleave JNK and NF-kB. EMBO J. 30, 221–231
Trends in Microbiology August 2013, Vol. 21, No. 8
85 Sham, H.P. et al. (2011) Attaching and effacing bacterial effector NleC
suppresses epithelial inflammatory responses by inhibiting NF-kB
and p38 mitogen-activated protein kinase activation. Infect. Immun.
79, 3552–3562
86 Rohde, J.R. et al. (2007) Type III secretion effectors of the IpaH family
are E3 ubiquitin ligases. Cell Host Microbe 1, 77–83
87 Li, H. et al. (2007) The phosphothreonine lyase activity of a bacterial
type III effector family. Science 315, 1000–1003
88 Arbibe, L. et al. (2007) An injected bacterial effector targets chromatin
access for transcription factor NF-kB to alter transcription of host
genes involved in immune responses. Nat. Immunol. 8, 47–56
89 Sauvonnet, N. et al. (2002) YopH prevents monocyte chemoattractant
protein 1 expression in macrophages and T-cell proliferation through
inactivation of the phosphatidylinositol 3-kinase pathway. Mol.
Microbiol. 45, 805–815
90 Orth, K. et al. (1999) Inhibition of the mitogen-activated protein kinase
kinase superfamily by a Yersinia effector. Science 285, 1920–1923
91 Bruno, V.M. et al. (2009) Salmonella Typhimurium type III secretion
effectors stimulate innate immune responses in cultured epithelial
cells. PLoS Pathog. 5, e1000538
92 Keestra, A.M. et al. (2013) Manipulation of small Rho GTPases is a
pathogen-induced process detected by NOD1. Nature 496, 233–237
93 Stecher, B. et al. (2007) Salmonella enterica serovar typhimurium
exploits inflammation to compete with the intestinal microbiota.
PLoS Biol. 5, 2177–2189
94 Lopez, C.A. et al. (2012) Phage-mediatedacquisition of a type III secreted
effector protein boosts growth of salmonella by nitrate respiration.
MBio 3, e00143–12, (http://dx.doi.org/10.1128/mBio.00143-12)
95 Winter, S.E. et al. (2010) Gut inflammation provides a respiratory
electron acceptor for Salmonella. Nature 467, 426–429
96 Raymond, B. et al. (2011) The WxxxE effector EspT triggers
expression of immune mediators in an Erk/JNK and NF-kB-
dependent manner. Cell. Microbiol. 13, 1881–1893
97 Fukazawa, A. et al. (2008) GEF-H1 mediated control of NOD1
dependent NF-kB activation by Shigella effectors. PLoS Pathog. 4,
e1000228
98 Keestra, A.M. et al. (2011) A Salmonella virulence factor activates the
NOD1/NOD2 signaling pathway. mBio 2, e00266–11, (http://
dx.doi.org/10.1128/mBio.00266-11)
99 Bergsbaken, T. et al. (2009) Pyroptosis: host cell death and
inflammation. Nat. Rev. Microbiol. 7, 99–109
100 Mariathasan, S. et al. (2004) Differential activation of the
inflammasome by caspase-1 adaptors ASC and Ipaf. Nature 430,
213–218
101 Muller, A.J. et al. (2009) The S. Typhimurium effector SopE induces
caspase-1 activation in stromal cells to initiate gut inflammation. Cell
Host Microbe 6, 125–136
102 Miao, E.A. et al. (2010) Innate immune detection of the type III
secretion apparatus through the NLRC4 inflammasome. Proc.
Natl. Acad. Sci. U.S.A. 107, 3076–3080
103 Perez-Lopez, A. et al. (2013) Salmonella downregulates Nod-like
receptor family CARD domain containing protein 4 expression to
promote its survival in B cells by preventing inflammasome
activation and cell death. J. Immunol. 190, 1201–1209
104 Hersh, D. et al. (1999) The Salmonella invasin SipB induces
macrophage apoptosis by binding to caspase-1. Proc. Natl. Acad.
Sci. U.S.A. 96, 2396–2401
105 Hilbi, H. et al. (1998) Shigella-induced apoptosis is dependent on
caspase-1 which binds to IpaB. J. Biol. Chem. 273, 32895–32900
106 Zhang, Y. et al. (2005) Inhibition of MAPK and NF-kB pathways is
necessary for rapid apoptosis in macrophages infected with Yersinia.
J. Immunol. 174, 7939–7949
107 Philip, N.H. and Brodsky, I.E. (2012) Cell death programs in Yersinia
immunity and pathogenesis. Front. Cell. Infect. Microbiol. 2, 149
108 Brodsky, I.E. et al. (2010) A Yersinia effector protein promotes
virulence by preventing inflammasome recognition of the type III
secretion system. Cell Host Microbe 7, 376–387
109 Schotte, P. et al. (2004) Targeting Rac1 by the Yersinia effector protein
YopE inhibits caspase-1-mediated maturation and release of
interleukin-1b. J. Biol. Chem. 279, 25134–25142
110 LaRock, C.N. and Cookson, B.T. (2012) The Yersinia virulence effector
YopM binds caspase-1 to arrest inflammasome assembly and
processing. Cell Host Microbe 12, 799–805
441