Published online: 15 december 2011 | doi: 10.1038/nchembio.741

bacteria and host interactions in the gut epithelial barrier
hiroshi ashida1, michinaga ogawa1, minsoo Kim2, hitomi mimuro1 & chihiro sasakawa1,2*
The gut mucosa acts as a barrier against microbial invaders, whereas resident commensal and foreign invading bacteria interact intimately with the gut epithelium and influence the host cellular and immune systems. The epithelial barrier serves as an infectious foothold for many bacterial pathogens and as an entry port for pathogens to disseminate into deeper tissues. Enteric bacterial pathogens can efficiently infect the gut mucosa using highly sophisticated virulence mechanisms that allow bacteria to circumvent the defense barriers in the gut. We provide an overview of the components of the mucosal barrier and discuss the bacterial stratagems that circumvent these barriers with particular emphasis on the roles of bacterial effector proteins.

© 2011 Nature America, Inc. All rights reserved.


he intestinal epithelium functions to gain nutrients, retain water and electrolytes and form an efficient barrier against foreign antigens and microbes. The intestinal mucosa not only is critical for intestinal homeostasis but also serves as an infectious foothold for the microbiota and invading pathogens. Therefore, the gut epithelium uses multiple defense mechanisms against microbes, including the luminal microbiota, a mucus layer, epithelial integrity, epithelial cell turnover and innate or acquired immune responses1. Despite the presence of these defensive systems, pathogenic bacteria can invade spaces that are usually devoid of microbes. Indeed, enteric bacterial pathogens such as Salmonella spp., Shigella spp., Vibrio cholerae, enteropathogenic Escherichia coli (EPEC), enterohemorrhagic E. coli (EHEC) and Yersinia can efficiently infect and multiply within the gut mucosa. These pathogens use a virulenceassociated protein (effector) injectisome, termed a type III secretion system (T3SS), that allows the bacteria to subvert the luminal microbiota, hijack host signaling pathways, modulate innate immune responses and circumvent innate defense barriers in the gut. We discuss the microbiota and the mucosal barrier as well as the epithelium itself and highlight bacterial stratagems to circumvent these barriers with particular emphasis on the roles of bacterial effector proteins (Table 1).

Bacterial pathogens have evolved highly sophisticated protein export systems that have been classified into seven types (types I–VII). Many Gram-negative bacterial pathogens have a T3SS, a type IV secretion system (T4SS) or both. T3SSs are evolutionarily and structurally related to flagellar export systems, whereas T4SSs are related to bacterial conjugation systems that translocate DNA. The T3SS consists of approximately 20 highly conserved proteins2 that form a multiprotein complex composed of the following distinctive parts: (i) a basal body, which is the channel spanning the bacterial membrane periplasm; (ii) a needle structure, which is the core T3SS projection that spans the bacterial membranes and the extracellular space; and (iii) a needle tip, which orchestrates the insertion of the translocon that links the needle to the host membrane (Fig. 1a). T3SS assembly is a tightly regulated and ordered process in which the basal body is formed in the inner and outer membranes before the needle structure is generated. When the needle is completely

Virulence strategies

formed, the needle tip is secreted and the assembly of the apparatus is completed. The T3SS is activated upon contact with host cells via the needle tip; translocators are then secreted by the T3SS and create a pore in the host cell membrane that provides access to the host cell cytosol. The temporal ordering of T3SS assembly and effector secretion is therefore essential for bacterial virulence (Fig. 1a). Bacterial pathogenesis that uses T3SSs to deliver a subset of effector proteins into host cells is dependent on T3SS activity. Consistent with this notion, inactivating the T3SS can almost completely abolish pathogenesis2. Therefore, activating T3SS at the appropriate site and time promotes successful bacterial infection. T3SS activity, as well as effector production and secretion, is controlled by transcriptional regulators in response to the intestinal environment. For example, in Shigella, T3SS activity is responsive to variations in oxygen concentrations3 (Fig. 1b). Oxygen gradients are present between the intestinal tissues and anaerobic intestinal lumen, and Shigella in the lumen sense environmental oxygen through an unidentified O2 sensor and transduce signals to Fnr, a global regulator that modulates anaerobic metabolism. Fnr represses the transcription of spa32 and spa33, whose products regulate T3SS needle length and effector secretion, respectively. Therefore, Fnr prevents the secretion of effector proteins and primes the extension of the T3SS needle until the bacteria directly contact the target epithelial cells. When Shigella reach a relatively oxygenized area above the intestinal epithelial cells, Fnr-dependent gene repression is silenced, the T3SS is activated and invasion proceeds3. T3SS activity in Salmonella enterica var. Typhimurium (S. Typhimurium) also responds to the environmental pH4,5 (Fig. 1c). Salmonella infect the intestinal mucosa using two distinct and sequential pathways: the Salmonella pathogenicity island 1 (SPI-1)dependent pathway, which is involved in bacterial invasion, and the SPI-2–dependent pathway, which is involved in bacterial intracellular survival. Salmonella infection of the gut triggers inflammation due to the delivery of effectors via T3SS-1 and T3SS-2 encoded by SPI-1 and SPI-2, respectively6. Salmonella enter intestinal epithelial cells and phagocytic cells where they survive and replicate in maturing Salmonella-containing vacuoles (SCVs), compartments required for bacterial replication, intracellular survival and systemic infection6. Once Salmonella invade and replicate within host cells, Toll-like receptors (TLRs) in the host recognize pathogen-associated


Department of Microbiology and Immunology, Institute of Medical Science, University of Tokyo, Tokyo, Japan. 2Department of Infectious Disease Control, International Research Center for Infectious Disease, Institute of Medical Science, University of Tokyo, Tokyo, Japan. *e-mail:
nature chemical biology | VOL 8 | JANUARY 2012 |


2a). Examining the role of microbiota in chemically induced colitis in germ-free mice containing selective gut microbiota (called gnotobiotic mice) showed that acetate production by commensal bacteria or acetate administration stimulated the G protein–coupled receptor 43 (GPR43).000 species. Once the translocator at the tip of T3SS-2 forms a membrane pore. For instance. germ-free mice have altered compositions of CD4+ T cells and IgA-producing B cells in the lamina propria. Once bacteria contact host cells via the needle tip and activate the T3SS. Comparing the transcriptional profiles of the intestinal epithelia of germ-free and conventional piglets similarly shows that conventional epithelia expresses proteins that contribute to epithelial turnover and are involved in the biosynthesis of mucin. an important component of the gut mucosal barrier. which encodes the rabbit homolog of LL-37 and enhances bactericidal activity15. from the lumen to the blood stream of the host12. and use this host response as a cue to induce the expression of bacterial virulence genes encoding the T3SS-2 component and to assemble it4 (Fig. Although T3SS-2 assembly is primed within the SCVs. Salmonella sense TLR-induced acidification within SCVs and induce the expression of the virulence gene locus SPI-2 as well as T3SS-2 assembly. Likewise. prevents bacterial infection by directly affecting virulence gene expression. T3SS-2 senses the neutral pH outside the SCVs (Fig. Under the anaerobic conditions of the intestinal lumen. rabbits that were pretreated with butyrate and infected with Shigella had reduced colonic inflammation and bacterial loads in the stool because butyrate upregulates the expression of the Cap18 gene. All rights reserved. which occurs as a result of TLRmediated innate immune responses. thereby triggering effector secretion. Bifidobacteria species that produce high concentrations of acetate as an end product of carbohydrate metabolism can prevent EHEC infection and the release of Shiga toxin. The mammalian intestine contains approximately 1014 commensal bacteria.nature. Fnr prevents the secretion of T3SS effectors by repressing the transcription of spa32 and spa33. epithelial proliferation and bacterial pathogenesis9 (Fig. These luminal commensal bacteria are a predominant part of the microbiota (hereafter we refer to commensal bacteria as the microbiota). Eubacterium rectale and Roseburia species. 1c). transduces a nature chemical biology | VOL 8 | JANUARY 2012 | www. these bacteria can directly or indirectly promote resistance to pathogenic bacterial . The effector proteins can pass through the needle and translocation pore into the host cell cytosol. are poorly developed. Block effector secretion 3. and they contribute to intestinal digestive function. two translocator proteins are secreted via the T3SS and form a pore in the host cell membrane. host immunity. Salmonella sense the acidification of the SCVs. which occurs at a two-fold faster rate in conventional mice than in germ-free mice10. In addition. which are aggregated lymphatic follicles. Both of these outputs are important for resistance to pathogen colonization11. which in turn allows epithelial proliferation and injury repair14. O2.1038/nchembio. propionate. which senses an elevated pH. molecular patterns (PAMPs).NaTurE chEmical biology doi: 10. (c) Salmonella use pH as a signal for T3SS-2 assembly and effector secretion within SCVs. pH. Short-chain fatty acids (SCFAs). the needle tip protein is secreted. which activate the innate immune response to limit and clear the infection. are produced as end metabolites by the microbiota and profoundly influence gut barrier function. thereby triggering the delivery of effector proteins into and across the vacuolar membrane. a needle structure and a needle tip. Assemble T3SS 2. Peyer’s patches. the underlying tissue of epithelial mucosa. which is most likely a ternary complex formed within the bacterial cytosol under acidic conditions. 2b).741 a Host cell membrane Needle tip Needle REVIEW ARTICLE E ector Pore Translocator dissociation signal to the regulatory complex. butyrate. such as acetate. for instance. resulting in prolonged bacterial survival and replication5 (Fig. an acetate chemoattractant receptor on immune cells that regulates inflammatory responses and allows the gnotobiotic mice to efficiently recover from colitis13. (b) Shigella modulate T3SS activity by sensing environmental O2 conditions. In the intestines of germ-free mice. Segmented filamentous bacteria (SFB). Although the precise mechanism of SFB-promoted TH17 differentiation 37 Outer membrane Periplasm Inner membrane Type III secretion system Base microbiota as a major luminal barrier E ector b Low O2 High Shigella Low oxygen Fnr Transcription Spa32 Spa33 No e ector secretion Lumen Epithelial cells High oxygen Fnr Transcription Spa32 Spa33 E ector secretion © 2011 Nature America. formate or butyrate. and in immune system priming. When the needle structure is complete.18 (Fig. 1c). influence epithelial metabolism and stimulate both epithelial cell proliferation and gut immunity as well as competing with enteric pathogens7–9. 1c). produced mainly by Fecalibacterium prausnitzii. Microbiota contribute to the development and function of the immune system. which contribute to resistance against colonization by pathogens17. After the T3SS-2 is assembled. virulence in the gut is believed to be regulated by similar mechanisms. adhere to the luminal surface of Peyer’s patches in the mouse intestine and strongly stimulate the differentiation of T helper 17 (TH17) cells in the lamina propria. However. Salmonella sense the neutral pH outside of the SCVs and induce the disassembly of the regulatory complex. bile salts or ions) of T3SS activity are highly conserved across many Gram-negative bacterial pathogens. Secrete effector E ector Figure 1 | T3SS is a delivery system for bacterial effectors. representing approximately 1. a crucial factor in lethal infection. Microbiota limit bacterial colonization and stimulate epithelial turnover. The induction of T-lymphocyte subsets is augmented by distinctive species of the luminal microbiota. effector secretion is prevented by a regulatory complex. Sense neutral pH 4. (a) The T3SS is composed of a basal body. Basolateral E ector c SCV PAMPs TLRs Acidification Virulence gene↑ E ector Neutral pH Acid pH E ector Regulatory complex E ector Disassembly regulatory complex E ector 1. upregulating the expression of the epithelial antimicrobial peptide LL-37 and providing energy for the colonic epithelium. compared to conventional mice and rats16. Inc. Because environmentally responsive regulators (responding to temperature. Thus. An unidentified pH sensor.

Microbiota help exclude pathogens by preventing colonization through enhancement of intestinal immune development and the production of SCFAs and bactericidal proteins.1038/nchembio. Some Clostridium species promote the release of the active form of TGF-b.741 b Epithelial cells IgA Microbiota Lumen Lamina propria Peyer’s patch M cells Antimicrobial peptide : Macrophage : Dendritic cells : T cells : B cells c Salmonella growth Microbiota H2S Thiosulfate Tetrathionate ROS Butyrate Lumen Stx2 EHEC Shigella SFB Adherence Clostridium Epithelial cells Salmonella Neutrophil Epithelial cells Epithelial cells Basolateral GPR43 Acetate GPR43 Chemoattractant Serum amyloid A Inflammation TGF-β ? Immune cells Inflammation TH17 cell accumulation Naive Treg cells Inflammation Bacterial pathogens © 2011 Nature America. which has been traditionally used to study and characterize bacterial pathogenesis20.23. Formate. which induces the differentiation of naive T cells into Treg cells19 (Fig. Typhimurium– infected mouse models demonstrated that virulent bacteria can outgrow the microbiota by delivering T3SS effectors to invade and multiply within host cells that trigger gut inflammation7. 2b). Salmonella’s ability to use tetrathionate as a terminal electron acceptor during respiration gives S. (a) SCFAs produced as end metabolites by the microbiota are important elements that prevent the colonization of bacterial pathogens. Although the composition of the microbiota in the small intestines or colon of untreated mice is markedly different. Inc. The mucus layer is largely composed of mucin.REVIEW ARTICLE a Bifidobacterirum Acetate Microbiota Microbiota Antimicrobial peptides NaTurE chEmical biology doi: 10. where the bacteria interact with the oligosaccharides of secreted mucin glycoproteins. Gut inflammation triggered by enteric pathogens disrupts the microbiota. leads to the downregulation of invasion genes.24. the tissue beneath the mucosal epithelium that contains various myeloid and lymphoid cells. Typhimurium induces inflammation that causes the production of reactive oxygen species that react with luminal thiosulphate to form tetrathionate. indicating that SCFA concentrations affect the preferential infection of the small intestine by Salmonella9. (c) S. where they stimulate the production of the active form of TGF-b. S. SFB adhere to the luminal surface of Peyer’s patches. some bacterial pathogens can circumvent these resistance mechanisms by delivering effectors and toxins that alter the microbiota composition and thus allow pathogenic bacteria to successfully colonize the intestinal epithelium. inflammatory mucin as a major epithelial cell surface barrier nature chemical biology | VOL 8 | JANUARY 2012 | www.22. mucin secretion can change in response to luminal conditions or bacterial infection. remains partly unclear. to form tetrathionate (S4O62–). SFB adherence to Peyer’s patches seems to induce serum amyloid A. Typhimurium induces inflammation that is accompanied by the production of reactive oxygen species and nitric oxide radicals. 2c). (b) The epithelial cell layer separates the gut lumen from the lamina propria. However. T cells and B cells. Although mucin is constitutively secreted at a basal level. which consists of membrane-bound mucin glycoproteins. whereas the inner layer is devoid of bacteria26. In the inner layer. Epithelial cells and Paneth cells secrete antimicrobial peptides that help prevent bacteria from penetrating the inner mucus layer. which is present at higher concentrations in the small intestine than in the colon.27. leads to the upregulation of invasion genes in Salmonella. Nevertheless. During infection. typhimurium a growth advantage over microbiota in the inflamed gut25 (Fig. thereby preventing inflammatory responses. intestinal immune molecules and invading pathogens is paramount in determining the conditions in the gut. In another example. These species can react with luminal thiosulfate (S2O32–). a 38 compound produced to detoxify the hydrogen sulfide (H2S) produced by colonic bacteria. The gut epithelium is covered by a thick mucus layer that acts as a frontline defense barrier against the microbiota and pathogenic bacteria26. Mucins are produced and secreted by goblet cells throughout the intestinal tract. Figure 2 | interaction between the microbiota and gut pathogens.nature. and TH17 cells differentiate in the lamina . and they help remove the gut contents and intruding microbes. S. allowing pathogens to outgrow the luminal microbiota. Studies with acute Citrobacter rodentium– or S. which promotes Treg cell differentiation and accumulation in the colonic lamina propria. The lamina propria contains a large number of immune cells and acts as an effector site for IgA production and T-cell responses. the colonization of germ-free mice with a defined mix of Clostridium strains revealed that some Clostridium species stimulate the accumulation of Foxp3+ T regulatory (Treg) cells in the colonic lamina propria. The impact of the microbiota on colonization resistance is further illustrated in an antibiotic-treated mouse model of infection. Typhimurium promotes its own outgrowth over the gut microbiota using an interesting mechanism. it was poorly understood how enteropathogenic bacteria circumvent colonization resistance mechanisms. 3a) and contains two sublayers: an outer loose mucus layer and an inner firmly adherent mucus layer. Luminal antigens are transported through M cells to the subepithelial dome of Peyer’s patches. resulting in an amelioration of colitis in chemically induced mouse models of the disease19 (Fig. which is present at higher concentrations in the colon than the small intestine. 2b).21. Bacterial colonization is therefore limited to the outer loose mucus layer. which contain dendritic cells. butyrate. the epithelial cell proximal surface is covered with glycocalyx. migration and proliferation. which contains various digestive enzymes and antimicrobial peptides as well as immunoglobulins (Fig. 3a). enteric bacterial pathogens have evolved mechanisms to circumvent this mucus barrier and directly access the epithelial surface. By contrast. Functional crosstalk among the luminal microbiota. Salmonella can use tetrathionate as a terminal respiratory electron acceptor. Mucin secretion by goblet cells is regulated by a variety of secretagogues.27 (Fig. All rights reserved. including microbial products. streptomycin treatment substantially decreases the luminal concentrations of SCFAs21. an apolipoprotein produced by the epithelial cells in Peyer’s patches that influences cell adhesion. The Clostridium species form a thick bacterial colonizing layer on the epithelium. Until recently.

signaling mediators. 3b). Epithelial cell-cell adherence is sustained by tight junctions (apical multiprotein complexes that form a selectively permeable seal between cells). Although the mechanism remains . which is a G protein-coupled receptor. 3b). the breakdown of tight junctions during bacterial infection results in gut barrier failure.741 a Bacterial pathogens Secreted mucin REVIEW ARTICLE b MUC1 H. Activation of this pathway. Some enteric pathogens deploy enzymes. adherence junctions (junctions subjacent to tight junctions that form a strong interaction with junctional molecules between cells). (b) Pathogenic bacterial infection causes Muc2 production through TLR-dependent or TLR-independent NF-kB pathways. During Pseudomonas aeruginosa infection of the colonic epithelium. which subsequently facilitates the translocation of bacteria and the luminal 39 epithelial integrity nature chemical biology | VOL 8 | JANUARY 2012 | www. The bacterial lipoteichoic acid activates platelet-activating factor receptor.1038/nchembio. 4a). approximately five-fold more H. (a) The mucus layer serves as a frontline defense against intruding bacterial pathogens. growth factors and infectious bacteria26. the interaction of pathogens with mucin can contribute to pathogen elimination due to rapid mucin secretion and mucus shedding. For instance. which degrade mucin oligosaccharides and reduce mucus viscosity and the release of antimicrobial peptides. or mucin-degrading enzymes39. C. For example. whereas Muc1-/. platelet-activating factor receptor. gastric epithelial cells can shed Muc1 in response to H. a zinc metalloprotease. Cell-surface mucins can limit the colonization of bacterial pathogens. such as Pic. inoculation of Muc1-/. Similarly. Hap. tight junctions are highly dynamic structures. preventing bacterial adhesion to the epithelial surface32 (Fig. Tight junctions are composed of zonula occludens (ZO-1 and ZO-2) and junctional adhesion molecules (JAM-1. gap junctions (paired connexin hemichannels) and desmosomes (adhesive junctions between cells). the recognition of PAMPs via TLRs and NOD-like receptors activates the downstream inflammatory signaling pathways that induce the transcription of mucin genes26. pylori colonize Muc1-/. jejuni uses Muc2 as an environmental cue to modulate the expression of genes involved in colonization and pathogenesis33. Disrupting flagella function reduces pathogenicity. In contrast.mice develop severe atrophic gastritis and lose gastric parietal cells31. hormones. Enteric bacterial pathogens have evolved mechanisms to sense and circumvent the mucus barrier and reach the epithelial cell surface. pathogens have developed specific mechanisms to penetrate the mucus barrier. StcE. a serine protease that degrades mucin (Table 1)35. aureus LTA PAFR G protein MUC Microbiota Outer mucus layer MUC Inner mucus layer MUC Antimicrobial peptides Immunoglobulin EGFR ADAM 10 Src Ras MAPK NF-κB MUC MUC Intestine Lamina propria Goblet cell Cellsurface mucin O-glycosylation Goblet cell Muc2 Intestine © 2011 Nature America. Thus. and the released Muc1 acts as a decoy that binds pathogens. composed of tight junctions and adherence junctions. infection with the Gram-positive bacteria Staphylococcus aureus induces the transcription of the Muc2 gene through a TLR-independent pathway29. The mucus layer acts as a protective barrier against colonization. a zinc metalloprotease that cleaves mucin-type O-glycosylated proteins (Table 1)36. Inc. lipopolysaccharide (LPS) induces Muc2 gene transcription by activating a TLR4-dependent pathway28 (Fig. pylori infection.38. claudin and occludin).NaTurE chEmical biology doi: 10. often termed leaky gut.nature. For example.mice than colonize wild-type mice. although the mucus layer prevents bacterial pathogens from accessing and breaching the epithelial lining. leading to the distension of transmembrane tight junction strands and increased paracellular permeability45. Thus. 3b). stimulates the RasMAPK-NF-kB pathway and activates mucin gene transcription29 (Fig. and they functionally segregate the apically expressed membrane proteins from those expressed on the basolateral membrane on polarized epithelial cells (Fig.27.mice (Muc1 encodes a cellsurface mucin) but not wild-type mice with Campylobacter jejuni results in rapid systemic infection30. aeruginosa TLR4 Pathogens Mucusdegradating enzyme Lumen MUC S. The flagella and chemotaxis systems are used by many enteric pathogens to traverse the mucus layer and access the epithelial cell surface. Treating epithelial monolayers with TNF-a or IL-1b increases tight junction permeability by stimulating transcription and activation of myosin light chain kinase (MLCK)42–44. Figure 3 | The mucus layer as a gut barrier. PAFR. whereas some bacterial pathogens have mucolytic enzyme activities that destroy these mucus layers. The apical junctions.27. Therefore.41. inflammatory cytokines can disrupt tight junctions and impair gut barrier integrity. and transduces signals to the epidermal growth factor receptor (EGFR) via ADAM10 metalloproteinase activity. and their permeability is regulated by several physiological and pathophysiological conditions. in turn. Intriguingly. The phosphorylation of MLC by MLCK stimulates perijunctional actomyosin contraction. pylori Bacterial pathogens Mucus layer LPS P. All rights reserved. a Hap homolog with metalloprotease activity that is distinct from Hap (Table 1)37. Cell-cell and cell–basement membrane interactions in the epithelium form a barrier that prevents bacteria from translocating to the subepithelial layer. highlighting the ability of bacterial motility to promote infection34. TagA. Wild-type mice develop only mild gastritis after H. However. pylori infection for 2 months. mediators. Mucin expression can also be controlled at the transcriptional level through several mechanisms. consist of transmembrane and cytoplasmic scaffolding proteins that associate with actin filaments and regulate epithelial paracellular permeability40.

CypD Bcl2 MAPKK PI3K/AKT Abcf2 p38. Activation of Rho GTPases by the S. SopB indirectly activates Rho GTPases. the activation or inactivation of one Rho GTPase can cause an imbalance in other networks. SopE2 Unknown Unknown AvrA SopB EspF VacA CagA Unknown Cif NleH IpaB 40 nature chemical biology | VOL 8 | JANUARY 2012 | www. Function Degradation of mucin Degradation of mucin Degradation of mucin Disrupts cell-cell junctions and cell polarity Disrupts cell-cell junctions Disregulates TJs Disrupts TJs Disrupts TJs Disrupts TJs Disregulates TJs Disrupts cytoskeletal integrity Disrupt tight junctions Induces mitochondrial dysfunction Inhibits mitochondrial dysfunction Inhibits cell death Inhibits cell death Induces apoptosis Induces apoptosis Increases proliferation Inhibits apoptosis and proliferation Increases proliferation Arrests cell cycle Inhibits apoptosis and proliferation Arrests cell cycle host target(s) Mucin Mucin Mucin ZO-1. pylori Regulation of epithelial cell turnover H. 4e). a function required for Rho GTPase activation. Inc. pylori Factor(s) Pic StcE Hap. 4c and Table 1)47. contents across the damaged epithelial lining and further promotes barrier damage and disease progression. Bacterial pathogens use tight junctions to facilitate their interactions with the epithelium and penetrate into deeper tissues. Rho GTPases play pivotal roles: RhoA triggers actomyosin assembly and contraction through Rho-associated kinase (ROCK)-mediated MLC phosphorylation. Typhimurium also delivers effectors that disrupt tight junctions57–59 (Fig. Map Table 1 | Selected examples of bacterial pathogens and gut defense system bacterial stratagems Disruption of mucus layer Breach cell-cell junction bacteria Shigella. which can upregulate RhoA and downregulate Cdc42. the intestinal barrier mediated by tight junctions was not compromised54. VopS (Table 1). rodentium lacking the lifA genes. which convert inactive GDP-bound Rho GTPases to the active GTP-bound forms and disrupt tight junction structure and function. T3SS effectors or toxins from enteric pathogens can subvert host signal pathways that regulate actin organization and tight junctions. tight junctions. C. resulting in actin disorganization and tight junction disruption41. Typhimurium can be blocked with an inhibitor of geranylgeranylation. SopE. Because signaling of Rho GTPases is tightly interconnected. EAEC EHEC V. This ultimately disrupts the integrity of the actin cytoskeleton in epithelial cells56 (Fig. Typhimurium effectors facilitate the uptake of bacteria by epithelial cells. C. CKIε NEDD8 BI-1 Mad2L2 reference(s) 35 36 37. NleA binds to and interferes with COPII-dependent protein trafficking. which leads to actin filament stabilization and tight junction modification (Fig. Tight junction disruption by S.78 81 82 89 87 86 91 96 90 VopS SipA. rodentium infection of murine intestinal epithelial cells results in diminished barrier function associated with disrupted tight junctions53. which regulates actin filaments. 4b). which encode lymphostatin. Because the components of tight junctions are constantly recycled between the plasma membrane and cytosol.46 (Fig. via the T3SS55. PAR1 Unknown MLCK.76 77. rodentium secretes a lymphocyte inhibitory factor called lymphostatin. whereas SopE and SopE2 act as GEFs that activate Rac1 and Cdc42 (ref. VopS catalyzes the AMPylation—the transfer of AMP from ATP to the threonine residues—of proteins such as Rac1. thus altering tight junction components and permeability52. pylori C.38 61–63 64 65 66 48 49–51 52 56 57 72 72 75.1038/nchembio. parahaemolyticus Salmonella Epithelial cell death and shedding Shigella Salmonella EPEC H. 4b). 4d). The gastrointestinal pathogen Vibrio parahaemolyticus delivers one effector that induces cell rounding. When mice were infected with C. Because tight junction components interact with the actin cytoskeleton to sustain the integrity of cell-cell adherence. Il1r1–/– or Il18–/– mice. which also affects tight junction structure and function. To this end. Bax Cyclin D1 MCL-1 b-catenin. For example. and the Rho GTPase pathway. Map and NleA (Fig. Cdc42 and RhoA via a phosphodiester bond56. TagA CagA VacA Urease Unknown EPEC Map EspM NleA V. gastrointestinal pathogens subvert several signal pathways that regulate tight junctions such as the protein kinase C pathway. regulating tight junction components is important to maintain epithelial integrity. Typhimurium SopE-mediated intestinal inflammation is abrogated in Casp-1–/–. Some effectors directly modulate Rho GTPases by acting as guanine exchange factors (GEFs). 6). S. and Rac1 and Cdc42 activate the PAK-LIMK-cofilin pathway. cholerae H. ROCK IL-1R1 Cdc42 RhoA COPII Rho GTPase Rho GTPase .nature. which modulates actin cytoskeleton rearrangement1. All rights reserved. indicating that Rho GTPase activation by these effectors is a cue for tight junction disruption57. EPEC disrupts tight junctions by altering actin cytoskeleton remodeling through the T3SS effectors EspM. SopB.741 activates Cdc42. S. indicating © 2011 Nature America. rodentium EPEC Shigella TJ. whereas EspM activates RhoA through its GEF activity and alters the localization of tight junctions48–51.REVIEW ARTICLE NaTurE chEmical biology doi: 10. thereby blocking Rho GTPase signaling and reducing actin cytoskeleton remodeling.

AMPylated Rho GTPase blocks signaling and reduces actin cytoskeleton remodeling. When these bacteria colonize the epithelium.62. promotes both epithelial cell apoptosis and shedding69. expose the basolateral cell surface and alter epithelial polarity to promote colonization and access the underlying tissues. epithelial cell death and shedding Gut epithelial cells undergo cell death and shedding under physiological and pathophysiological conditions. JAM. pylori TJ ZO-1 CagA open Par1 EPEC C. they induce the demise of host cells to facilitate egress from their niches1. where they disrupt tight junctions and adherence junctions60. adherence junction. AJ.70. (d) V.NaTurE chEmical biology doi: 10.and cyclophilin D-dependent process. Some bacterial pathogens deliver effectors that modulate Rho GTPases. Although the mechanism of tight junction disruption during H. which are composed of finger-like projections that protrude from the epithelial lining of the intestinal wall. Some enteric pathogens can either induce or prevent cell death to gain a survival advantage in the gut71. However. Tight junctions form a semipermeable barrier that restricts the paracellular passage of water and nutrients and maintains cell polarity. allowing the bacteria to disrupt epithelial tight junctions. CagA triggers the mislocalization of ZO-1 from tight junctions to the bacterial attachment sites61. parahaemolyticus delivers VopS. 72). pylori upregulates the phosphorylation of IL-1 receptor type I. pylori CagA and VacA (Table 1) can be delivered into the gastric epithelium.1038/nchembio. implying that the pathogen can suppress cell death at the initial stage of infection73. proliferation and turnover. Typhimurium disrupt tight junctions by activating the Rho GTPase. Typhimurium manipulate actin filaments and intestinal inflammation. (e) The SipA. Cell death signaling becomes dominant through a BNIP3. SopE and SopE2 effectors of S. which has AMPylation activity.74. The Salmonella AvrA and SopB effectors counteract epithelial cell death. disrupting this balance can lead to pathological conditions. a balance among epithelial cell proliferation. parahaemolyticus e TJ Salmonella TJ open SipA SopB SopE SopE Rho GTPase SopE2 Rho GTPase Casp-1 IL-1β Actin remodeling TJ open Map VopS AMP Rho GTPase AMP EspM RhoA lifA Cdc42 Rho GTPase Actin remodeling © 2011 Nature . SopB. death and exfoliation is required to maintain homeostasis and epithelial integrity. During bacterial infection. pylori urease (Table 1) stimulates MLCK and ROCK activation through unknown mechanisms. All rights reserved. As many as ~3% of total epithelial cells from the villi. which occurs only after prolonged exposure to the pathogens. In addition.68 (Fig. H. which targets ZO-1 and the PAR1 (also called MARK) polarity-regulating kinase and attenuates gastric epithelial barrier function by altering epithelial cell polarity61–63 (Fig. For instance. pylori infection is still speculative. which subsequently activates ROCK and disrupts tight junctions through a process that is independent of CagA and VacA66. obtain nutrients and access the underlying tissues. AvrA. and targets Rho GTPase. Inc. rodentium Actin remodeling d V. thereby disrupting epithelial integrity63. differentiation. Typhimurium infection of porcine intestines or Salmonella typhimurium serovar Dublin infection of human intestinal epithelial cells induces caspase-3–dependent and caspase-3–independent cell death. an acetyltransferase that 41 nature chemical biology | VOL 8 | JANUARY 2012 | www. S. H. a cytokine that is elevated in inflammatory bowel disease.nature. 4c). epithelial cell death is a host defense response that eliminates damaged cells as well as pathogens and ultimately emits alarm signals that activate the innate immune system. VacA also helps open tight junctions64. In addition. migration. pylori effectors can disrupt gut integrity via multiple mechanisms. leading to the dysregulation of tight junctions65. are extruded in the lumen in the human and mouse intestine67. inflammatory diseases such as ulcerative colitis increase tight junction permeability as well as epithelial apoptosis. Adhesion junctions are composed of members of the cadherin family. In mice. Therefore. CagA binds to PAR1 and inhibits its activity. they first prevent cell death to preserve their replicative foothold. Tight junctions are composed of ZO-1. Intestinal epithelial cells are tightly linked to neighboring cells via cell-cell junctions and form defensive barriers to prevent bacterial intrusion. thereby inducing cell rounding. (b) PKC and Rho GTPases trigger tight junction disruption by modulating the actin cytoskeleton. whereas later. many bacterial pathogens can breach epithelial integrity. H. (a) A schematic of junction proteins in polarized epithelial cells. (c) Bacterial pathogens target tight junctions and breach epithelial integrity to promote colonization. Tra cking COPII NleA Cell rounding Inflammation IL-18 Figure 4 | bacterial pathogens breach epithelial integrity. Shigella invasion of epithelial cells results in mitochondrial dysfunction—mediated necrosis-like cell death. 5a). which leads to mitochondrial permeability and cell death72. pylori delivers CagA via a T4SS. These studies suggest that S. H. that caspase-1 activation and cytokine production are involved in the disruption of tight junctions59. The phosphorylation of MLC by MLCK induces the contraction of perijunctional actomyosin and breaches the tight junction barrier. TNF-a. H. and disrupting these allows the bacteria to directly access basolaterally localized receptors such as b1-integrin or c-Met. but the cell death response is antagonized until the bacteria have fully replicated through activation of prosurvival pathway via the recognition of PAMPs (peptidoglycan) by Nod1 (ref.741 a Apical Lumen REVIEW ARTICLE b TJ open Actin remodeling TJ JAM ZO-1 Occludin Claudin Actomyosin open P MLC P P TJ AJ TJ P Cofilin LIMK PAK PKC MLCK Desmosome Intermediate filaments E-cadherin AJ Gap junctions Actin cytoskeleton P ROCK Rac1 Cdc42 Basolateral Rho GTPase RhoA c H. claudin and occludin. H. pylori targets cell-cell junctions. Thus.

pylori. disruption of epithelial intercellular junctions. such as H.80. The epithelial cells that line the gastrointestinal lumen are constantly renewed through a process in which stem cells generated in the crypts migrate to the tip of the villi and ultimately peel off into the lumen. allowing Salmonella to counteract epithelial cell apoptosis77. C. The gut epithelium undergoes continuous self-renewal to maintain tissue homeostasis and eliminate damaged cells.741 b EPEC Salmonella H. an inositol phosphatase. Shigella can manipulate epithelial cell turnover by directly accessing cryptic epithelial progenitors. Colonization of the gastric superficial epithelium by H. However.1038/nchembio. All rights reserved.REVIEW ARTICLE a Shedding cell NaTurE chEmical biology doi: 10. Some bacterial pathogens.78. rodentium stimulates the proliferation of cryptic stem cells. Inc. Some EPEC strains deliver Cif (Table 1).nature. thereby inducing apoptosis81. resulting in cytochrome c release and apoptosis79. helps dampen inflammatory and cell death responses75. in which p38 activation and Bax oligomerization have a key role. pylori can attenuate apoptosis of maturated gastric pit cells. whereby they breach the epithelial barrier. As a result. specifically. SopB. modulates MAPPKs and inhibits the JNK signaling pathway. 5b). pylori colonization causes apoptosis of gastric pit cells by inducing oxidative stress. their death facilitates bacterial egress and spread to other host tissues. IpaB was shown to negatively regulate cell-cycle progression by directly binding to Mad2L2. an antiapoptosis factor. EspF interacts with Abcf2. Bacterial pathogens can also manipulate neddylation (attachment of the ubiquitin-like protein NEDD8) to regulate epithelial turnover and promote infection90–92. In HeLa cells and rabbit ileal-loop infection . which plays a critical role in cell survival and proliferation. There is a balance between the elimination of damaged cells and the generation of new cells from a stem cell population in the intestinal crypt (Fig. H. (a) The gut epithelium undergoes continuous self-renewal to maintain tissue homeostasis and eliminate damaged cells. counteract rapid epithelial turnover and maintain epithelial cells as a replicative niche. CagA-mediated upregulation of these transcription factors results in the production of cyclin D1. pylori CagA hijacks multiple cell signaling pathways to promote persistent colonization of the gastric epithelium by activating transcription factors such as nuclear factor of activated T cells. access the underlying tissues and obtain nutrients. and reduces its expression within the mitochondria. a cell-cycle regulator and the enhanced survival of epithelial cells88. leading to hyperplasia and increased crypt length85. 5b). which blocks the cell nature chemical biology | VOL 8 | JANUARY 2012 | www. to promote bacterial colonization89. Epithelial cell death at the early stage of infection is predominantly a consequence of host defense mechanisms. some pathogens elicit epithelial cell death. NF-kB or T-cell factor–lymphoid enhancer factor (TCF-LEF)60. which are normally shed every 2–3 days in a gerbil model of infection86. pylori infection augmented epithelial expression of the prosurvival factor Erk and the antiapoptotic protein Mcl-1 in a CagA-dependent manner. rodentium stimulates b-catenin 42 epithelial turnover signaling via upregulating casein kinase Iε (CKIε) production. enhancing the bacterial load in the gerbil stomach86 (Fig. H. When epithelial cell apoptosis was stimulated with etoposide. whereas some bacterial pathogens deploy countermeasures against rapid epithelial turnover to prolong their survival. VacA-mediated translocation of Bax oligomers into the mitochondria in response to H.87. Figure 5 | bacterial countermeasures against epithelial cell death and turnover. activates the serine-threonine kinase Akt. Aside from physiological renewal. and these effectors have an impact on a variety of host cell-signaling pathways that are involved in cytoskeletal rearrangement. Selected effectors are shown. an effector with a papain-like hydrolytic fold that includes a cysteine-histidine-glutamine catalytic triad. In mice. H. the bacteria deliver the IpaB effector (Table 1) via a T3SS into progenitors.84. pylori infection results in decreased mitochondrial membrane potential and apoptosis82. Shigella or EPEC. epithelial turnover can accelerate (or decelerate) in response to changes in the luminal environment. epithelial cell death and immune modulation47. pylori causes an imbalance between epithelial cell proliferation and apoptosis86. 5a). pylori Epithelial cell Villus CagA AvrA Ac MAPKK Cif Gln40→Glu40 deamidation NEDD8 Cullin-RING ubiquitin ligase ERK Migration Crypt Di erentiation Lamina propria Progenitor cell Stem cell Paneth cell Ac MAPKK MCL1 Cytochrome c JNK Cell-cycle arrest Casp-3 Apoptosis Proliferation and turnover © 2011 Nature America. These studies show that increased epithelial cell turnover in response to bacterial pathogens is a host defense mechanism that eliminates infected cells. H. EspF disrupts the mitochondrial membrane potential.76 (Fig. Epithelial turnover of the Drosophila melanogaster gut accelerates in response to infection with Erwinia carotovora. limits persistent bacterial colonization and maintains tissue homeostasis. an anaphase-promoting complex inhibitor. (b) Epithelial cell death and turnover are highly dynamic responses to bacterial infection and limit persistent bacterial colonization. Pseudomonas spp. After the bacteria fully propagate within epithelial cells. C. serum response factor. or Serratia marcescens83. EPEC delivers a subset of effectors (Table 1) via the T3SS.

Enteric pathogens can deploy countermeasures to prevent rapid turnover of epithelial cells in order to maintain epithelial cells as a replicative niche. 12. USA 103. This report shows the benefit of intestinal inflammation in promoting the colonization of bacterial pathogens. S. 5. E. and several small-molecule compounds (both synthetic compounds and natural products) have been identified as T3SS inhibitors. We envision that an understanding of the mechanisms by which bacteria disrupt the mutualism between the mucosa and microbiota as well as promote infection of the gut epithelium will provide avenues to conclusions and perspectives 1. 2691–2702 (2009). Garner. et al. 215 (2007). a group of salicylidene acylhydrazides inhibits the T3SS activity of many bacterial pathogens by blocking the assembly of the T3SS needle complex. Liu. B. T.P. Science 331. J. 16. Bacteriol. & Finlay. Enteric pathogen exploitation of the microbiotagenerated nutrient environment of the gut. 675–688 (2011). The development of various new technologies. Opin.V. B. 23. K. Independent of its kinase activity. During EPEC infection. Microbiol. et al. H..J.L. 9. et al. 5. Gaboriau-Routhiau.D. C. The key role of segmented filamentous bacteria in the coordinated maturation of gut helper T cell responses.J. Garner. Lawley. Cicconi. Stecher. 92–98 (2011). Acad. microbiota composition and gut homeostasis. NleH inhibits several cellular responses associated with apoptosis.M.. Y. & Wolf-Watz. Stecher. One relatively new approach. excessive host inflammatory responses. McGhie. Nat. & Macpherson. Microbiol. Salmonella enterica serovar typhimurium exploits inflammation to compete with the intestinal microbiota. D. C. 2. B. Nat. Immunity 31. C. Opin. Mechanisms controlling pathogen colonization of the gut. Bacterial interactions with the host epithelium. Cell Host Microbe 8. Immun.C. & Altier. Shifting the balance: antibiotic effects on host-microbiota mutualism. McGourty. Transcriptome profiling of the small intestinal epithelium in germfree versus conventional piglets. Rev. 14.M. & Holden. rodentium colonization sites in the intestine95. Humphreys. N. Typhimurium-induced host inflammation changes the composition and suppresses the growth of the microbiota. Bifidobacteria can protect from enteropathogenic infection through production of acetate. Hooper. Siegel. Immun. 24. K. In recent years.97. S. Immune adaptations that maintain homeostasis with the intestinal microbiota.B. Inhibition of these enzymes may induce formation of actin stress fibers and contribute to cell-cycle arrest90. Enteric bacterial pathogens have highly evolved mechanisms to shift the balance of host-microbiota mutualism for the pathogen’s benefit. From the gut to the peripheral tissues: the multiple effects of butyrate. Galán. Raqib. Kim.. Antibiotics have been extensively used to selectively target bacterial pathogens to treat and prevent many infectious diseases96. Curr.. 1040–1043 (2010). et al. 9178–9183 (2006). Russell. 82–91 (2011). including elevation of cytoplasmic Ca2+ concentrations. Snellen. 159–169 (2010). Transit time of epithelial cells in the small intestines of germfree mice and ex-germfree mice associated with indigenous microorganisms. P.. allows us to comprehensively understand how the gut commensal (and pathogenic) bacteria establish replicative niches and how the host immune system responds to bacterial infection. J.J.D. 233–243 (2011). 43 nature chemical biology | VOL 8 | JANUARY 2012 | www. 543–547 (2011). 117–124 (2009). Microbiol. & Hardt. Nutr. et al. K. Natl. Opin. Gut inflammation provides a respiratory electron acceptor for Salmonella. The authors showed that S. Infect. D. 10. D. These findings suggest that epithelial turnover changes in response to bacterial infection to limit persistent bacterial colonization. references Mutualism between the gut mucosa and microbiota is the most important factor in sustaining gut homeostasis. W.nature. et al. Nature 469. which sustains the replicative niche. In this regard. 95). This study shows that acetate produced by Bifidobacteria species prevents lethal infection and epithelial cell death induced by EHEC O157.. 13.. et al. Microbiol. 22. T3SS inhibitors have been recently considered new drugs. X. Rev. © 2011 Nature America. For example.E. Maslowski. 677–689 (2009). Salmonella takes control: effector-driven manipulation of the host. 426–429 (2010).R.. thereby promoting the emergence of antibiotic-resistant bacteria. Unsworth. Ivanov. 2177–2189 (2007).J. Formate acts as a diffusible signal to induce Salmonella invasion. et al. 119–129 (2007). Nature 461. S. 996–1001 (1981). L.E. 19. 5b). Immunol. Arpaia. Science 328. A. Cell 144. 337–341 (2011).D. 25. systems biology. Brawn. Induction of intestinal Th17 cells by segmented filamentous bacteria. K. circumventing the innate barriers of the gut and modulating mucosal innate immune responses.NaTurE chEmical biology doi: 10. Suyemoto. TLR signaling is required for Salmonella typhimurium virulence. Perturbation of the small intestine microbial ecology by streptomycin alters pathology in a Salmonella enterica serovar typhimurium murine model of infection. 6. 9. Huang. I. Nature 444.. Savage. 3. thereby overcoming colonization resistance.. Appl. Sci. pH sensing by intracellular Salmonella induces effector translocation. NleH inhibited procaspase-3 cleavage at C. K. bioinformatics. V. BMC Genomics 8. Many enteric Gram-negative pathogens deploy T3SS effectors that promote the survival and colonization of these pathogens within the gut by disrupting microbiota-mediated colonization resistance. & Koronakis. 17. Regulation of inflammatory responses by gut microbiota and chemoattractant receptor GPR43. Fukuda. 1282–1286 (2009). Cell Host Microbe 2. the effector NleH (Table 1) inhibits apoptosis by binding to the endoplasmic reticulum six-transmembrane protein Bax inhibitor (BI-1). In a mouse model of C. 20–35 (2010). 4233–4241 (2008).D. we need to generate new drugs that target bacterial pathogenesis rather than kill pathogenic bacteria96. 366–384 (2010). 567–573 (2006). 77. Protein delivery into eukaryotic cells by type III secretion machines.B. 8.97. Lupp. & Finlay. 190. J. Host-mediated inflammation disrupts the intestinal microbiota and promotes the overgrowth of Enterobacteriaceae. & Whitt. Res. et al. Environ. S. nuclear condensation and activation of caspase-3 (ref. PLoS Biol. antibiotics also disrupt the composition of the microbiota. Hume. 23. REVIEW ARTICLE develop new therapeutics that control microbial infection. Induction of colonic regulatory T cells by indigenous Clostridium species. M.. T3SS effectors are bacterial executioners that can directly or indirectly modify the target host proteins and subvert host cellular and immune functions. Microbiol. R. Cell 139. New insights are providing a blueprint for the dynamic networks among the commensal (and pathogenic) microbes and the gut immune system in condition of both health and disease. 18. B. C. animal and other eukaryotic model systems will help elucidate the unidentified mechanisms of effector proteins and provide insights that can be used to develop new antimicrobial drugs and therapeutics as alternatives to antibiotics.1038/nchembio.741 cycle by causing both G1-S and G2-M arrest93. Winter. Inc. 20. M. 4. Infect. Rev. Although the exact role of NleH in EPEC infection remains partly speculative. rodentium infection. Willing.C. 485–498 (2009). Keeney. thereby preventing T3SS effector secretion and virulence98–100. 76. 14. B. K. there has been an explosion of new analytical tools that help to identify the microbial and host factors affecting the mutualistic relationship between the microbes and gut. inhibiting the activity of the NEDD8-conjugated Cullin-RING E3 ubiquitin ligases90 (Fig. et al. Improved outcome in shigellosis associated with butyrate induction of an endogenous peptide antibiotic. Nature 465. et al. 14. et al.E. 42. 7. This study reports that Clostridium species induce regulatory T cells and maintain immunological homeostasis via stimulating matrix metalloprotease-TGF-b signaling.W. However. V.D.94. P.. J. . 11.E. et al. Curr. Curr. D. et al. often leaving an imprint on the composition of the microbiota after the antibiotic treatment has been discontinued. Host transmission of Salmonella enterica serovar Typhimurium is controlled by virulence factors and indigenous intestinal microbiota. 355–358 (2010).I. 403–416 (2008). et al. Cif selectively binds to NEDD8 and deamidates Gln40. Marteyn. Proc. M. it is likely that NleH facilitates EPEC pathogenesis by reducing enterocyte loss.E. 10.M. Chowdhury. Modulation of Shigella virulence in response to available oxygen in vivo. Atarashi. All rights reserved. L. Nature 467. 21. et al. B. 15. Thus. Guilloteau. Yu.

73. Piguet. Chem. B. & Florin. pylori colonization. Microbiol.B. M.C. Carneiro. Halladin. Microbiol. Science 300. & Buret. 12. 48. McGuckin. Biochim. (in the press). et al.A. Ye. T. PLoS Pathog. NY Acad. EspM inhibits pedestal formation by enterohaemorrhagic 51. 44 This study.M. & Montrose. Ramos. M. 237–243 (2005).. L.W. e1000538 (2009). Infect. 509–517 (2004). 29..M. et al. 64. & Chadee... Tumor necrosis factor-induced long myosin light chain kinase transcription is regulated by differentiation-dependent signaling events. This review highlights the role of tight junctions as a component of the epithelial barrier and describes how bacterial pathogens target and alter tight junctions during infection. 16339–16344 (2005). Innate Immun.H. 12. M. 26–38 (2010). a secreted protease of Shigella flexneri and enteroaggregative Escherichia coli. 1210–1218 (2007). Cell. Invest. 54. & DeVinney.. Am. 799–809 (2009). Acad. 43. 76. lessons from toxins. A. ZVAD-fmk. Rev. I. Schofield. Flynn.L. 75.A. Saadat.. S. Gastroenterology 140.H. & Basbaum. Dharmani. Mucin dynamics and enteric pathogens. Ann. et al.. 4131–4138 (1993). T. 59. SopE. Haemagglutinin/protease expression and mucin gel penetration in El Tor biotype Vibrio cholerae. 654–664 (2010). 5718–5723 (1998). 5. et al. & Welch. 12. J. Babbin. 28. J. Alto. Cell. C. 31–41 (2010). 795–802 (2008). & Finlay. Mantle. Interferon-g and tumor necrosis factor-a synergize to induce intestinal epithelial barrier dysfunction by up-regulating myosin light chain kinase expression. 236–246 (2009). Cell adhesion: the molecular basis of tissue architecture and morphogenesis. et al. 32. 39. Interleukin-1 receptor phosphorylation activates Rho kinase to disrupt human gastric tight junctional claudin-4 during Helicobacter pylori infection. R. Donati. Acta 1788. 61. Immunol. Med. 67. Schauser. Tompkins. B.. J.. Activation of NF-kB via a Src-dependent Ras-MAPKpp90rsk pathway is required for Pseudomonas aeruginosa-induced mucin overproduction in epithelial cells. S. 71. Helicobacter pylori CagA induces a transition from polarized to invasive phenotypes in MDCK cells. Typhimurium use tetrathionate. 1295–1303 (2005).A. 137. 4. Mechanism of IL-1b-induced increase in intestinal epithelial tight junction permeability. Arbeloa. Southward.. 79. USA 102. 8.J. et al. MUC1 cell surface mucin is a critical element of the mucosal barrier to infection.. 124..L. & Galán. 123–136 (2009). L. 73. & Kagnoff. Microbiol. Ashida. & Finlay. Multifaceted role of Rho. Du. Muc1 mucin limits both Helicobacter pylori colonization of the murine gastric mucosa and associated gastritis. D. Li. 102. 44. et al. Pathobiology of the intestinal epithelial barrier. R.A. Helicobacter pylori dysregulation of gastric epithelial tight junctions by urease-mediated myosin II activation. et al. AMPylation of Rho GTPases by Vibrio VopS disrupts effector binding and downstream signaling. e1000617 (2009).. 1430–1434 (2003). SopE2 and SipA disrupt tight junction structure and function. R.. V.M. 1946–1957 (2006). 58. Nusrat. J. McGuckin. 813–820 (1998). 9. Selective increase of the permeability of polarized epithelial cell monolayers by Helicobacter pylori vacuolating toxin.R. et al.A. et al. Menard. Tight junctions as targets of infectious agents. Identification of a bacterial type III effector family with G protein mimicry functions. et al.R. B. & Larsson. Subversion of actin dynamics by EspM effectors of attaching and effacing bacterial pathogens. This study shows that VopS has AMPylation activity and modifies conserved threonine residues in Rho GTPase. Gerner-Schmidt. Papini..F. coli and disrupts the architecture of a polarized epithelial monolayer. Microbiology 157. J. D. A. Salmonella Typhimurium type III secretion effectors stimulate innate immune responses in cultured epithelial cells. 70. J. 46.G..J. Microbiology 149.B. Buti. Covacci. S. Amieva..V. 45. It also describes the interaction between bacterial pathogens and the mucus layer. S.REVIEW ARTICLE NaTurE chEmical biology doi: 10. Microbiol. MUC1 limits Helicobacter pylori infection both by steric hindrance and by acting as a releasable decoy. R. Grys. Gastroenterology 136. C.W.M. Pathol.. Srivastava. Cell Host Microbe 5. Tu. The StcE protease contributes to intimate adherence of enterohemorrhagic Escherichia coli O157:H7 to host cells. 345–357 (1996). 30. Microbiol. Microbiol.A. J. Immun. H. R. The bacterial virulence factor lymphostatin compromises intestinal epithelial barrier function by modulating rho GTPases. Virulence mechanisms and persistence strategies of the human gastric pathogen Helicobacter pylori. & Barazzone. M. 41.E.. N. Guiney. 269–272 (2009)..D. Infect.. Eslava. Paesold. Cell Biol. Infect. et al. A. Med. Trends Microbiol. Marchiando.L. 495–500 (1999).. & Ma. Brown. Sci. Szabady. 1. Vesin. J. Shigella induces mitochondrial dysfunction and cell death in nonmyeloid cells. The S. All rights reserved. F. K. M. 52. Czeczulin. USA 95. 516–525 (2011). Sci. Campylobacter jejuni response to human mucin MUC2: modulation of colonization and pathogenicity determinants. as an electron acceptor and gains a growth advantage to overcome the host microbiota. F. Pathol. Casselli. J.Y. Invest. Siegel. shows the benefits of intestinal inflammation during S. Bacterial flagellins: mediators of pathogenicity and host immune responses in mucosa. 1273–1281 (1990). Y. Characterization of the human long myosin light chain kinase promoter.M. Acad. Microbiol.D.M. Growth in and breakdown of purified rabbit small intestinal mucin by Yersinia enterocolitica. Thanabalasuriar. F. & Buret. A. M. Sci. Microbiol. 489–505 (2010). Mechanisms of epithelial cell shedding in the Mammalian intestine and maintenance of barrier function.K. 27.A. TagA is a secreted protease of Vibrio cholerae that specifically cleaves mucin glycoproteins. This review explores the role of mucin as a barrier to bacterial infection. 61. D. H. K. J. Immunol. Pathog. Pham.741 Escherichia coli and enteropathogenic E. O’Connor. et al. 37. 23. et al.K..A. 55. I. A.. Cell. V. Yanta. Lynch. J. 281. T. Silva. 69. A. Inc. Cell. Guttman. Biol. E. Intestinal mucosal barrier function in health and disease.. Graham.K. 60. Bagnoli. N. T. Vibrio parahaemolyticus inhibition of Rho family GTPase activation requires a functional chromosome I type III secretion system. & Geny. Natl. 125–136 (2009).A. 67. 692–703 (2010). 76. 1429–1441 (2008). 26205–26215 (2006). Natl. E. in addition to ref. M. B. Proc. A. The authors found that mucin acts as decoy that is released from the epithelial surface in response to bacterial binding. Sasaki. M. 1347–1357 (2009).A. 47. 74. Lathem. V. 8. et al. 5653–5661 (2008). The bacterial virulence factor NleA is required for the disruption of intestinal tight junctions by enteropathogenic Escherichia coli. Genes in the Salmonella pathogenicity island 2 and the Salmonella virulence plasmid are essential for Salmonella-induced apoptosis in intestinal epithelial cells. Curr. Cell.N. Henderson.J. & Haas. et al. & Sirard. Cell death and infection: a double-edged sword for host and pathogen survival. Cell Biol. M. Jones. et al. & Welch. Characterization of pic.B. Madara. Nature 447. 63. M. Wang. & Amieva. 68. & Mendz. 1883–1891 (2003). 34.E. L. 98–104 (2008).. Science 323. A. 56.G. Cell. 174. D. 41–46 (2002). Boyle. & Klapproth. et al. 1165. 33. Guan. 53. Top. 65. Lab. Jones.. Nat. M. 135–142 (2009). Gumbiner. Bruno. Lindén.P. Fischer.W. 57. Simovitch. Eckmann. 42. Typhimurium effector SopE induces caspase-1 activation in stromal cells to initiate gut inflammation. Al-Sadi.R. P. B.R. J. 117. Popoff. 35. Microb. Dokladny. K. The epithelial barrier is maintained by in vivo tight junction expansion during pathologic intestinal epithelial shedding. DNA fragmentation and apoptotic morphology. 57. P. Nat. Pathol. R. EspM2 is a RhoA guanine nucleotide exchange factor. K. Watson. Salmonella AvrA coordinates suppression of host immune and apoptotic defenses via JNK pathway blockade. B.. nature chemical biology | VOL 8 | JANUARY 2012 | www. Nat. J. 9. Rumbo.. 5. Programmed cell death and cell extrusion in rat duodenum: a study of expression and activation of caspase-3 in relation to C-jun phosphorylation.L. Biochim. 49.. & Nataro. Cell Host Microbe 20. C. H. F.B. Disruption of the epithelial apical-junctional complex by Helicobacter pylori CagA. Clin. 5587–5596 (1999). et al. Role of intestinal mucins in innate host defense mechanisms against pathogens. 166. W. Cell. PLoS Pathog. 409–419 (2005).com/naturechemicalbiology . 123–135 (2009). Selective inhibition of type III secretion activated signaling 26. M. Tight junctional disruption and apoptosis in an in vitro model of Citrobacter rodentium infection. & Finlay. T. Rac. Prassl.H. T. Proc. Noriega. Am..V.M. A. Warner-Lambert/Parke-Davis Award lecture.C. C. J. Lapointe. A. 12. 36.K. J. Lemjabbar. M. 797–812 (2009). J. Müller.nature. J. M. Molecular mechanisms of Escherichia coli pathogenicity. 50. & Benitez. Arbeloa. J. C. 10. Salmonella enterica serovar Typhimurium effectors SopB. 40. thereby disrupting Rho GTPase signaling and regulating actin cytoskeleton remodeling. Croxen. Immunol. 12.. R. et al. 45. 133–145 (2006). Duckworth. Typhimurium infection.G.L. Y.A. Acta 1788. P. McAuley.J. J.M. Q. 233–244 (2008). 31. 2202–2211 (2008).E. 8. Nat. 38. Histochem. Am. Infect. 2313–2324 (2007). A.. Cell Host Microbe 3. Jones. Turner. Helicobacter pylori CagA targets PAR1/MARK kinase to disrupt epithelial cell polarity. 771–781 (2002). 832–841 (2009). which is produced as a result of inflammation. L. P. et al. Lindén. W. J.1038/nchembio. M. Immun.C. B. This study shows the importance of MUC1 in limiting H. Rev. Platelet-activating factor receptor and ADAM10 mediate responses to Staphylococcus aureus in epithelial cells. G. Sutton. Biophys. TNF-induced enterocyte apoptosis and detachment in mice: induction of caspases and prevention by a caspase inhibitor. Cdc42 and Ras in intercellular junctions. Rev. 129–171 (2009). Biophys. M.. Microbiol..F. G. 265–278 (2011). Microbiol. McGuckin. Invest. Clin. Immun.F. K. Wroblewski. Cell 84.. Gastroenterology 133. Microbiol. 1208–1218 (2011). 330–333 (2007). A. 337. & Rombough. Cell 124. W. Immun. 180. N.I. 62. L. et al. thereby preventing prolonged infection. 66. © 2011 Nature America. C. L. J.L..R. C. Kissoon-Singh. 72. Yarbrough. J.

Veenendaal. 2496–2508 (2008).). 87. Helicobacter pylori dampens gut epithelial self-renewal by inhibiting apoptosis. G. PLoS Pathog. 89. 401. 85. Knodler. Sellin. A. A bacterial effector targets Mad2L2..K. 3716–3721 (2009). a Grant-in-Aid for Scientific Research (S) (20229006 to C. Science 329.NaTurE chEmical biology doi: 10.B.T. Wang. Cell Stem Cell 4.B. 84. H. et al. 79. 315. 92.. 280. J. L. Lo. and 23790472 to H. 268–274 (2010). Nordfelth. a Grant-in-Aid for Scientific Research (B) (23390102 to H. et al. a bacterial strategy to enhance colonization of the stomach. 563–570 (2009).A. B. Agents Chemother. C. B.H. 1–16 (2006). Sundin. 397–405 (2008).H.. et al. N. 294. a Grant-in-Aid for Young Scientists (A) (23689027 to M. et al. M. Small-molecule inhibitors specifically targeting type III secretion. Molecular mechanisms of epithelial-barrier disruption by Helicobacter pylori. Med. Enteropathogenic Escherichia coli EspF is targeted to mitochondria and is required to initiate the mitochondrial death pathway. 611–623 (2007). REVIEW ARTICLE Commun. Acad. A bacterial type III effector family uses the papain-like hydrolytic activity to arrest the host cell cycle. & Finlay.W. Singh. et al. 3129–3134 (2010). et al. Part of this work was supported by grants from the Naito Foundation (to H. USA 106.1038/nchembio. 93. A. et al. 6. et al. Proc. PLoS Pathog. G. Homeostasis in infected epithelia: stem cells take the lead.S. Immun. 9. Pathogenic bacteria target NEDD8-conjugated cullins to hijack host-cell signaling pathways. Iwai. and a grant from the Japan Initiative for Global Research Network on Infectious Diseases to C. 7. A. Enteropathogenic Escherichia coli effector EspF interacts with host protein Abcf2. Drugs R D. Biol. Yu. Correspondence and requests for materials should be addressed to C. Biol. & Umar. J. Bacterial cyclomodulin Cif blocks the host cell cycle by stabilizing the cyclin-dependent kinase inhibitors p21 and p27. to modulate host cell cycling. J. Apidianakis. Salicylidene acylhydrazides that affect type III protein secretion in Salmonella enterica serovar typhimurium. et al. The bacterial effector Cif interferes with SCF ubiquitin ligase function by inhibiting deneddylation of Cullin1. Microbiol. C. H. Nougayrède. J. & Donnenberg.. Hemrajani. M. A. C. 1215–1218 (2010).B. Natl..nature. Physiol. Marra. Y.K. Cell 130. Cell. Chem. Jiang. 2554–2566 (2011). 100. Gastrointest. Microbiol. Nagai. a Grant-in-Aid for Young Scientists (B) (23790471 to M. an APC inhibitor. 16. Immun. Liver Physiol. Inc. 86. G635–G647 (2008).nature. 77.741 by the Salmonella effector AvrA. Am. H. S. Morikawa.M. 94. J. & Donnenberg. C.. 10.J.. Nougayrède. acknowledgments © 2011 Nature America.A. A. Mimuro. A.). 82. S. e1000595 (2009). & Sasakawa. & Perrimon. reprints/index.S.. Levy. R.).P. All rights reserved.S.C. resulting in cell cycle arrest. 191. J. 91. & Backert. & Blocker. Samba-Louaka. Cui. & Elofsson. S122–S129 (2004). 97–109 (2009). 79. 96.). Cell Res. 3104–3114 (2005). Nat. H. Science and Technology. 9058–9064 (2005). NleH effectors interact with Bax inhibitor-1 to block apoptosis during enteropathogenic Escherichia coli infection. Sports. This work was supported by Grant-in-Aid for Specially Promoted Research (23000012 to C. 98. Sci. 83. J. Q. 51. Small-molecule type III secretion system inhibitors block assembly of the Shigella type III secreton. Mechanisms for Helicobacter pylori CagA-induced cyclin D1 expression that affect cell cycle. Cell Host Microbe 6. Kauppi. Sci. 90. Targeting virulence for antibacterial chemotherapy: identifying and characterising virulence factors for lead discovery.). Negrea. Antibacterial resistance worldwide: causes.P. Yao. The Salmonella effector protein SopB protects epithelial cells from apoptosis by sustained activation of Akt. Reprints and permissions information is available online at http://www.. & Ip.S. Acad. Norberg. Cell Host Microbe 2. Natl. & Marshall. from the Ministry of Education. & Steele-Mortimer. competing financial interests additional information The authors declare no competing financial interests. 78. Trends Microbiol. O. Protective role of Akt2 in Salmonella enterica serovar typhimurium-induced 45 . Glutamine deamidation and dysfunction of ubiquitin/NEDD8 induced by a bacterial effector family. Microbiol. Biophys. Microbiol. M. 5. b-Catenin stabilization imparts crypt progenitor phenotype to hyperproliferating colonic epithelia. S. Targeting of enteropathogenic Escherichia coli EspF to host mitochondria is essential for bacterial pathogenesis: critical role of the 16th leucine residue in EspF. 2867–2876 (2007). B. Res. Exp. challenges and responses. 49–61 (2009). Bacteriol. H. 8. 80.O. USA 107.A. A. Y. et al. M. A.R.. This study reveals that Cif deaminates NEDD8 and interferes with its function. Y. Tissue damage-induced intestinal stem cell division in Drosophila. Cell. Amcheslavsky. 301–307 (2009). 97.M. et al. Differential regulation of ERK1/2 and p38 MAP kinases in VacA-induced apoptosis of gastric epithelial cells. 1097–1111 (2004). Biochem.M. Wolf-Watz. Chang.S.. 250–263 (2007). Finlay.S. Cell. a Grant-in-Aid for Challenging Exploratory Research (23659220 to H.html.).B. Abe.. Foster. 6. 2998–3011 (2005). H. Ki. e1001128 (2010). 10. Cell. nature chemical biology | VOL 8 | JANUARY 2012 | www. 280.J. W. S. Y. 99.). Kum.. 73. Pitsouli. J. P. 88. Jubelin. Proc. Chem. 680–693 (2007). 95. Infect. Wessler. 81. B. Antimicrob. 1740–1752 (2006). Culture. J. Infect.