Functionally Essential Interaction Between Yersinia YscO and the T3S4 Domain of YscP

Overview

Journal J Bacteriol

Publisher American Society for Microbiology

Specialty Microbiology

Date 2013 Aug 13

PMID 23935040

Citations 6

Authors

Romila Mukerjea

Partho Ghosh

Affiliations

Soon will be listed here.

Abstract

The type III secretion (T3S) system is essential to the virulence of a large number of Gram-negative bacterial pathogens, including Yersinia. YscO is required for T3S in Yersinia and is known to interact with several other T3S proteins, including the chaperone SycD and the needle length regulator YscP. To define which interactions of YscO are required for T3S, we pursued model-guided mutagenesis: three conserved and surface-exposed regions of modeled YscO were targeted for multiple alanine substitutions. Most of the mutations abrogated T3S and did so in a recessive manner, consistent with a loss of function. Both functional and nonfunctional YscO mutant proteins interacted with SycD, indicating that the mutations had not affected protein stability. Likewise, both functional and nonfunctional versions of YscO were exclusively intrabacterial. Functional and nonfunctional versions of YscO were, however, distinguishable with respect to interaction with YscP. This interaction was observed only for wild-type YscO and a T3S-proficient mutant of YscO but not for the several T3S-deficient mutants of YscO. Evidence is presented that the YscO-YscP interaction is direct and that the type III secretion substrate specificity switch (T3S4) domain of YscP is sufficient for this interaction. These results provide evidence that the interaction of YscO with YscP, and in particular the T3S4 domain of YscP, is essential to type III secretion.

Citing Articles

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References

Lupas A, Van Dyke M, Stock J . Predicting coiled coils from protein sequences. Science. 1991; 252(5009):1162-4. DOI: 10.1126/science.252.5009.1162. View

Cornelis G . The type III secretion injectisome. Nat Rev Microbiol. 2006; 4(11):811-25. DOI: 10.1038/nrmicro1526. View

Bzymek K, Hamaoka B, Ghosh P . Two translation products of Yersinia yscQ assemble to form a complex essential to type III secretion. Biochemistry. 2012; 51(8):1669-77. PMC: 3289748. DOI: 10.1021/bi201792p. View

Journet L, Agrain C, Broz P, Cornelis G . The needle length of bacterial injectisomes is determined by a molecular ruler. Science. 2003; 302(5651):1757-60. DOI: 10.1126/science.1091422. View

Evans L, Hughes C . Selective binding of virulence type III export chaperones by FliJ escort orthologues InvI and YscO. FEMS Microbiol Lett. 2009; 293(2):292-7. PMC: 3500872. DOI: 10.1111/j.1574-6968.2009.01535.x. View

Neyt C, Cornelis G . Role of SycD, the chaperone of the Yersinia Yop translocators YopB and YopD. Mol Microbiol. 1999; 31(1):143-56. DOI: 10.1046/j.1365-2958.1999.01154.x. View

Lorenzini E, Singer A, Singh B, Lam R, Skarina T, Chirgadze N . Structure and protein-protein interaction studies on Chlamydia trachomatis protein CT670 (YscO Homolog). J Bacteriol. 2010; 192(11):2746-56. PMC: 2876502. DOI: 10.1128/JB.01479-09. View

Riordan K, Schneewind O . YscU cleavage and the assembly of Yersinia type III secretion machine complexes. Mol Microbiol. 2008; 68(6):1485-501. PMC: 3538813. DOI: 10.1111/j.1365-2958.2008.06247.x. View

Arnold K, Bordoli L, Kopp J, Schwede T . The SWISS-MODEL workspace: a web-based environment for protein structure homology modelling. Bioinformatics. 2005; 22(2):195-201. DOI: 10.1093/bioinformatics/bti770. View

10.

Payne P, Straley S . YscP of Yersinia pestis is a secreted component of the Yop secretion system. J Bacteriol. 1999; 181(9):2852-62. PMC: 93729. DOI: 10.1128/JB.181.9.2852-2862.1999. View

11.

Gouet P, Robert X, Courcelle E . ESPript/ENDscript: Extracting and rendering sequence and 3D information from atomic structures of proteins. Nucleic Acids Res. 2003; 31(13):3320-3. PMC: 168963. DOI: 10.1093/nar/gkg556. View

12.

Woestyn S, Allaoui A, Wattiau P, Cornelis G . YscN, the putative energizer of the Yersinia Yop secretion machinery. J Bacteriol. 1994; 176(6):1561-9. PMC: 205240. DOI: 10.1128/jb.176.6.1561-1569.1994. View

13.

Evans L, Stafford G, Ahmed S, Fraser G, Hughes C . An escort mechanism for cycling of export chaperones during flagellum assembly. Proc Natl Acad Sci U S A. 2006; 103(46):17474-9. PMC: 1859953. DOI: 10.1073/pnas.0605197103. View

14.

Edqvist P, Olsson J, Lavander M, Sundberg L, Forsberg A, Wolf-Watz H . YscP and YscU regulate substrate specificity of the Yersinia type III secretion system. J Bacteriol. 2003; 185(7):2259-66. PMC: 151483. DOI: 10.1128/JB.185.7.2259-2266.2003. View

15.

LARKIN M, Blackshields G, Brown N, Chenna R, McGettigan P, McWilliam H . Clustal W and Clustal X version 2.0. Bioinformatics. 2007; 23(21):2947-8. DOI: 10.1093/bioinformatics/btm404. View

16.

Allaoui A, Woestyn S, Sluiters C, Cornelis G . YscU, a Yersinia enterocolitica inner membrane protein involved in Yop secretion. J Bacteriol. 1994; 176(15):4534-42. PMC: 196272. DOI: 10.1128/jb.176.15.4534-4542.1994. View

17.

Diepold A, Wiesand U, Amstutz M, Cornelis G . Assembly of the Yersinia injectisome: the missing pieces. Mol Microbiol. 2012; 85(5):878-92. DOI: 10.1111/j.1365-2958.2012.08146.x. View

18.

Riordan K, Sorg J, Berube B, Schneewind O . Impassable YscP substrates and their impact on the Yersinia enterocolitica type III secretion pathway. J Bacteriol. 2008; 190(18):6204-16. PMC: 2546781. DOI: 10.1128/JB.00467-08. View

19.

Lathem W, Price P, Miller V, Goldman W . A plasminogen-activating protease specifically controls the development of primary pneumonic plague. Science. 2007; 315(5811):509-13. DOI: 10.1126/science.1137195. View

20.

Sorg I, Wagner S, Amstutz M, Muller S, Broz P, Lussi Y . YscU recognizes translocators as export substrates of the Yersinia injectisome. EMBO J. 2007; 26(12):3015-24. PMC: 1894769. DOI: 10.1038/sj.emboj.7601731. View