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SycE allows secretion of YopE-DHFR hybrids by the Yersinia enterocolitica type III Ysc system

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Feldman, MF Muller, S Wuest, E Cornelis, Guy

The Ysc type III secretion system allows Yersinia enterocolitica to translocate virulence proteins, called Yop effectors, into the cytosol of eukaryotic cells. Some of the Yop effectors possess an individual chaperone called a Syc protein. The first 15 amino acids of the YopE effector constitute a secretion signal that is sufficient to promote secretion of several reporter proteins. Residues 15-50 of YopE comprise the minimal binding domain for the SycE chaperone. In this study, we investigated the secretion by the Ysc system of several YopE-DHFR hybrid proteins with different folding properties, and evaluated the role of SycE, the cognate chaperone of YopE, in this context. We have analysed the secretion of hybrids containing 16 (YopE(16)), 52 (YopE(52)) and 80 (the complete region covered by the chaperone, YopE(80)) amino acids of YopE or full-length YopE (YopE(FL)) with wild-type DHFR and two mutants with altered folding properties. The hybrids containing DHFRDelta77, the mutant whose folding properties are the most drastically affected, could be secreted in all the conditions tested, even in the absence of the chaperone SycE. In contrast, DHFRwt could only be secreted fused to the first 52 amino acids of YopE, and its secretion was strictly dependent on SycE. The hybrids YopE(80)-DHFRwt and YopE(FL)-DHFRwt were not secreted. YopE(FL)-DHFRwt completely jammed the channel in an SycE-dependent fashion. Our experiments indicate that, in order to be secreted, proteins must be unfolded or only partially folded, and that TSS chaperones could keep their substrates in a secretion-competent conformation, probably by preventing their folding. In addition, they show that the secretion apparatus can reject folded proteins if they are not deeply engaged into the injectisome.

Document type Article de périodique (Journal article) – Article de recherche
Publication date 2002
Language Anglais
Journal information "Molecular Microbiology" - Vol. 46, no. 4, p. 1183-1197 (2002)
Peer reviewed yes
Publisher Blackwell Publishing Ltd (Oxford)
issn 0950-382X
e-issn 1365-2958
Publication status Publié
Affiliation UCL
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Bibliographic reference Feldman, MF ; Muller, S ; Wuest, E ; Cornelis, Guy. SycE allows secretion of YopE-DHFR hybrids by the Yersinia enterocolitica type III Ysc system. In: Molecular Microbiology, Vol. 46, no. 4, p. 1183-1197 (2002)
Permanent URL http://hdl.handle.net/2078.1/41507