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Volume 165, Issue 1

Editor's Choice SrcA is a chaperone for the Salmonella SPI-2 type three secretion system effector SteD Open Access

Abstract

Effector proteins of type three secretion systems (T3SS) often require cytosolic chaperones for their stabilization, to interact with the secretion machinery and to enable effector delivery into host cells. We found that deletion of srcA, previously shown to encode a chaperone for the Salmonella pathogenicity island 2 (SPI-2) T3SS effectors SseL and PipB2, prevented the reduction of mature Major Histocompatibility Complex class II (mMHCII) from the surface of antigen-presenting cells during Salmonella infection. This activity was shown previously to be caused by the SPI-2 T3SS effector SteD. Since srcA and steD are located in the same operon on the Salmonella chromosome, this suggested that the srcA phenotype might be due to an indirect effect on SteD. We found that SrcA is not translocated by the SPI-2 T3SS but interacts directly and forms a stable complex with SteD in bacteria with a 2 : 1 stoichiometry. We found that SrcA was not required for SPI-2 T3SS-dependent, neutral pH-induced secretion of either SseL or PipB2 but was essential for secretion of SteD. SrcA therefore functions as a chaperone for SteD, explaining its requirement for the reduction in surface levels of mMHCII.

  • Received:
  • Accepted:
  • Published Online:
Keyword(s): chaperone , effector , Salmonella and type III secretion system
© 2019 The Authors
  • This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
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2018-11-20
2024-04-18
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References

  1. Portaliou AG, Tsolis KC, Loos MS, Zorzini V, Economou A. Type III secretion: building and operating a remarkable nanomachine. Trends Biochem Sci 2016; 41:175–189 [View Article][PubMed]
     [Google Scholar]
  2. Bennett JC, Hughes C. From flagellum assembly to virulence: the extended family of type III export chaperones. Trends Microbiol 2000; 8:202–204 [View Article][PubMed]
     [Google Scholar]
  3. Parsot C, Hamiaux C, Page AL. The various and varying roles of specific chaperones in type III secretion systems. Curr Opin Microbiol 2003; 6:7–14 [View Article][PubMed]
     [Google Scholar]
  4. Stebbins CE, Galán JE. Priming virulence factors for delivery into the host. Nat Rev Mol Cell Biol 2003; 4:738–744 [View Article][PubMed]
     [Google Scholar]
  5. Fu Y, Galán JE. Identification of a specific chaperone for SptP, a substrate of the centisome 63 type III secretion system of Salmonella typhimurium. J Bacteriol 1998; 180:3393–3399[PubMed]
     [Google Scholar]
  6. Ogawa M, Suzuki T, Tatsuno I, Abe H, Sasakawa C. IcsB, secreted via the type III secretion system, is chaperoned by IpgA and required at the post-invasion stage of Shigella pathogenicity. Mol Microbiol 2003; 48:913–931 [View Article][PubMed]
     [Google Scholar]
  7. Thomas NA, Deng W, Puente JL, Frey EA, Yip CK et al. CesT is a multi-effector chaperone and recruitment factor required for the efficient type III secretion of both LEE- and non-LEE-encoded effectors of enteropathogenic Escherichia coli. Mol Microbiol 2005; 57:1762–1779 [View Article][PubMed]
     [Google Scholar]
  8. Jennings E, Thurston TLM, Holden DW. Salmonella SPI-2 type III secretion system effectors: molecular mechanisms and physiological consequences. Cell Host Microbe 2017; 22:217–231 [View Article][PubMed]
     [Google Scholar]
  9. Cheminay C, Möhlenbrink A, Hensel M. Intracellular Salmonella inhibit antigen presentation by dendritic cells. J Immunol 2005; 174:2892–2899 [View Article][PubMed]
     [Google Scholar]
  10. Lapaque N, Hutchinson JL, Jones DC, Méresse S, Holden DW et al. Salmonella regulates polyubiquitination and surface expression of MHC class II antigens. Proc Natl Acad Sci USA 2009; 106:14052–14057 [View Article][PubMed]
     [Google Scholar]
  11. Bayer-Santos E, Durkin CH, Rigano LA, Kupz A, Alix E et al. The Salmonella effector SteD mediates MARCH8-dependent ubiquitination of MHC II molecules and inhibits T cell activation. Cell Host Microbe 2016; 20:584–595 [View Article][PubMed]
     [Google Scholar]
  12. Cooper CA, Zhang K, Andres SN, Fang Y, Kaniuk NA et al. Structural and biochemical characterization of SrcA, a multi-cargo type III secretion chaperone in Salmonella required for pathogenic association with a host. PLoS Pathog 2010; 6:e1000751 [View Article][PubMed]
     [Google Scholar]
  13. Yu XJ, McGourty K, Liu M, Unsworth KE, Holden DW. pH sensing by intracellular Salmonella induces effector translocation. Science 2010; 328:1040–1043 [View Article][PubMed]
     [Google Scholar]
  14. Scheich C, Kümmel D, Soumailakakis D, Heinemann U, Büssow K. Vectors for co-expression of an unrestricted number of proteins. Nucleic Acids Res 2007; 35:e43 [View Article][PubMed]
     [Google Scholar]
  15. Studier FW. Protein production by auto-induction in high density shaking cultures. Protein Expr Purif 2005; 41:207–234 [View Article][PubMed]
     [Google Scholar]
  16. Sagné C, Isambert MF, Henry JP, Gasnier B. SDS-resistant aggregation of membrane proteins: application to the purification of the vesicular monoamine transporter. Biochem J 1996; 316:825–831 [View Article][PubMed]
     [Google Scholar]
  17. Srikumar S, Kröger C, Hébrard M, Colgan A, Owen SV et al. RNA-seq Brings New Insights to the Intra-Macrophage Transcriptome of Salmonella Typhimurium. PLoS Pathog 2015; 11:e1005262 [View Article][PubMed]
     [Google Scholar]
  18. Paul P, van den Hoorn T, Jongsma ML, Bakker MJ, Hengeveld R et al. A Genome-wide multidimensional RNAi screen reveals pathways controlling MHC class II antigen presentation. Cell 2011; 145:268–283 [View Article][PubMed]
     [Google Scholar]
  19. Bijlmakers MJ, Benaroch P, Ploegh HL. Assembly of HLA DR1 molecules translated in vitro: binding of peptide in the endoplasmic reticulum precludes association with invariant chain. EMBO J 1994; 13:2699–2707 [View Article][PubMed]
     [Google Scholar]
  20. Lloyd SA, Norman M, Rosqvist R, Wolf-Watz H. Yersinia YopE is targeted for type III secretion by N-terminal, not mRNA, signals. Mol Microbiol 2001; 39:520–532 [View Article][PubMed]
     [Google Scholar]
  21. Shi Y, Mowery RA, Ashley J, Hentz M, Ramirez AJ et al. Abnormal SDS-PAGE migration of cytosolic proteins can identify domains and mechanisms that control surfactant binding. Protein Sci 2012; 21:1197–1209 [View Article][PubMed]
     [Google Scholar]
  22. Evdokimov AG, Tropea JE, Routzahn KM, Copeland TD, Waugh DS. Structure of the N-terminal domain of Yersinia pestis YopH at 2.0 A resolution. Acta Crystallogr D Biol Crystallogr 2001; 57:793–799 [View Article][PubMed]
     [Google Scholar]
  23. Stebbins CE, Galán JE. Maintenance of an unfolded polypeptide by a cognate chaperone in bacterial type III secretion. Nature 2001; 414:77–81 [View Article][PubMed]
     [Google Scholar]
  24. Allison SE, Tuinema BR, Everson ES, Sugiman-Marangos S, Zhang K et al. Identification of the docking site between a type III secretion system ATPase and a chaperone for effector cargo. J Biol Chem 2014; 289:23734–23744 [View Article][PubMed]
     [Google Scholar]
  25. Birtalan SC, Phillips RM, Ghosh P. Three-dimensional secretion signals in chaperone-effector complexes of bacterial pathogens. Mol Cell 2002; 9:971–980 [View Article][PubMed]
     [Google Scholar]
  26. Abe A, de Grado M, Pfuetzner RA, Sánchez-Sanmartín C, Devinney R et al. Enteropathogenic Escherichia coli translocated intimin receptor, Tir, requires a specific chaperone for stable secretion. Mol Microbiol 1999; 33:1162–1175 [View Article][PubMed]
     [Google Scholar]
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SrcA is a chaperone for the Salmonella SPI-2 type three secretion system effector SteD
Microbiology 165, 15 (2019); https://doi.org/10.1099/mic.0.000732
/content/journal/micro/10.1099/mic.0.000732
/content/journal/micro/10.1099/mic.0.000732
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