Three-dimensional Structure of a Macromolecular Assembly that Regulates Type III Secretion in Yersinia pestis

Schubot, Florian D.; Jackson, Michael; Penrose, Kerri J.; Cherry, Scott; Tropea, Joseph E.; Plano, Gregory V.; Waugh, David S.

doi:10.1016/j.jmb.2004.12.036

Cited by 99 publications

(171 citation statements)

References 52 publications

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“…Work from several groups further suggests that the C-terminally located effector domains are folded and catalytically active in the presence of their respective chaperones (9,14,21,24). Only recently, the structure of the secretable regulatory TTSS component YopN from Yersinia in complex with the heterodimeric chaperone SycN/YscB was presented (17). This structure unambiguously confirms that the influence of chaperone binding on folding of the secretion substrate is confined to the chaperone-binding site and does not extend globally.…”

mentioning

confidence: 80%

“…This hypothesis is supported by the impressive conservation of folds among class I chaperones of TTSSs given their low sequence similarity. Further, it is supported by the structural conservation of the mode of effector/chaperone interaction, which is not based on a high degree of sequence similarity (9,13,16,17). This functional conservation of chaperone activity is underscored by the interchangeability of effector/chaperone pairs among TTSSs of different species (5,42).…”

Section: Discussionmentioning

confidence: 99%

“…Chaperones of class II assist the TTSS translocators, and the flagellar TTSS chaperones constitute class III according to this classification. Crystal structures of representatives of class IA (Salmonella SicP (13) and SigE (14); Yersinia SycE (15), SycH (16), and SycN/YscB (17); Escherichia coli CesT (14); Pseudomonas syringae AvrPphF ORF1 (18)), class IB (Shigella flexneri Spa15 (19)), and class III (Aquifex aeolicus FliS (20)) support this classification. The class IA chaperones SicP, SigE, SycE, CesT, and AvrPphF ORF1, although not similar on a sequence level, all form homodimers and share a common fold.…”

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confidence: 83%

“…ComplexesWork from several groups suggests that the influence of class I TTSS chaperones on the folding of their substrates is restricted to the binding site generally encompassing about 50 -100 amino acid residues (9,14,17,21,24). To understand the functioning of these chaperones, high resolution structural data on complete type III effectors in complex with their respective chaperones are required.…”

Section: Co-expression and Purification Of Yopt/syctmentioning

confidence: 99%

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Crystal Structure of the Yersinia enterocolitica Type III Secretion Chaperone SycT

Locher

Lehnert

Krauss

et al. 2005

Journal of Biological Chemistry

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Several Gram-negative pathogens deploy type III secretion systems (TTSSs) as molecular syringes to inject effector proteins into host cells. Prior to secretion, some of these effectors are accompanied by specific type III secretion chaperones. The Yersinia enterocolitica TTSS chaperone SycT escorts the effector YopT, a cysteine protease that inactivates the small GTPase RhoA of targeted host cells. We solved the crystal structure of SycT at 2.5 Å resolution. Despite limited sequence similarity among TTSS chaperones, the SycT structure revealed a global fold similar to that exhibited by other structurally solved TTSS chaperones. The dimerization domain of SycT, however, differed from that of all other known TTSS chaperone structures. Thus, the dimerization domain of TTSS chaperones does not likely serve as a general recognition pattern for downstream processing of effector/chaperone complexes. Yersinia Yop effectors are bound to their specific Syc chaperones close to the Yop N termini, distinct from their catalytic domains. Here, we showed that the catalytically inactive YopT C139S is reduced in its ability to bind SycT, suggesting an ancillary interaction between YopT and SycT. This interaction could maintain the protease inactive prior to secretion or could influence the secretion competence and folding of YopT.

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mentioning

confidence: 80%

Section: Discussionmentioning

confidence: 99%

mentioning

confidence: 83%

Section: Co-expression and Purification Of Yopt/syctmentioning

confidence: 99%

See 2 more Smart Citations

Crystal Structure of the Yersinia enterocolitica Type III Secretion Chaperone SycT

Locher

Lehnert

Krauss

et al. 2005

Journal of Biological Chemistry

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“…The C-terminal domains in the InvE/MxiC/CopN/SepL/ YopN-TyeA family of proteins (TyeA in the case of the YopN-TyeA heterodimer) are important for regulating the secretion of translocases or effector proteins (Archuleta et al, 2011;Deane et al, 2008;Schubot et al, 2005;Wang et al, 2008). To investigate whether the C terminus of InvE is involved in the regulation of SipB secretion, we constructed plasmids that encoded FLAG-tagged InvE containing full-length or C-terminal-truncated forms and introduced them into the DinvE mutant.…”

Section: Resultsmentioning

confidence: 99%

Molecular characterization of the InvE regulator in the secretion of type III secretion translocases in Salmonella enterica serovar Typhimurium

et al. 2013

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The type III secretion systems (T3SSs) are exploited by many Gram-negative pathogenic bacteria to deliver a set of effector proteins into the host cytosol during cell entry. The T3SS of Salmonella enterica serovar Typhimurium is composed of more than 20 proteins that constitute the membrane-associated base, the needle and the tip complex at the distal end of the T3SS needle. Membrane docking and piercing between the T3SS and host cells is followed by the secretion of effector proteins. Therefore, a secretion hierarchy among the substrates of the T3SS is required. The secretion of the pore-forming translocase proteins SipB, SipC and SipD is controlled by the T3SS regulator protein, InvE. During an attempt to identify the regions of InvE that are involved in T3SS regulation, it was observed that the secretion of SipB, SipC and SipD was inhibited when the C-terminal 52 amino acids were removed from InvE. In addition, InvE derivatives lacking the Nterminal 30 and 100 residues were unable to secrete translocases into the culture medium. Interestingly, in the absence of the N-terminal 180 residues of InvE, SipD is unstable, resulting in the hypersecretion of SipB. We also found that both the type III secretion signals of SipB and SptP were functionally interchangeable with the first 30 amino acids of InvE, which could allow the secretion of a reporter protein. These results indicate that InvE may have two functional domains responsible for regulating the secretion of translocases: an N-terminal secretion signal and a Cterminal regulatory domain.

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Novel insights into the mechanism of SepL‐mediated control of effector secretion in enteropathogenic Escherichia coli

et al. 2017

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Type three secretion systems (T3SSs) are virulence determinants employed by several pathogenic bacteria as molecular syringes to inject effector proteins into host cells. Diarrhea‐producing enteropathogenic Escherichia coli (EPEC) uses a T3SS to colonize the intestinal tract. T3S is a highly coordinated process that ensures hierarchical delivery of three classes of substrates: early (inner rod and needle subunits), middle (translocators), and late (effectors). Translocation of effectors is triggered upon host‐cell contact in response to different environmental cues, such as calcium levels. The T3S substrate specificity switch from middle to late substrates in EPEC is regulated by the SepL and SepD proteins, which interact with each other and form a trimeric complex with the chaperone CesL. In this study, we investigated the link between calcium concentration and secretion regulation by the gatekeeper SepL. We found that calcium depletion promotes late substrate secretion in a translocon‐independent manner. Furthermore, the stability, formation, and subcellular localization of the SepL/SepD/CesL regulatory complex were not affected by the absence of calcium. In addition, we demonstrate that SepL interacts in a calcium‐independent manner with the major export gate component EscV, which in turn interacts with both middle and late secretion substrates, providing a docking site for T3S. These results suggest that EscV serves as a binding platform for both the SepL regulatory protein and secreted substrates during the ordered assembly of the T3SS.

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Three-dimensional Structure of a Macromolecular Assembly that Regulates Type III Secretion in Yersinia pestis

Cited by 99 publications

References 52 publications

Crystal Structure of the Yersinia enterocolitica Type III Secretion Chaperone SycT

Crystal Structure of the Yersinia enterocolitica Type III Secretion Chaperone SycT

Molecular characterization of the InvE regulator in the secretion of type III secretion translocases in Salmonella enterica serovar Typhimurium

Novel insights into the mechanism of SepL‐mediated control of effector secretion in enteropathogenic Escherichia coli

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