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Volume 150, Issue 7

The SPI2-encoded SseA chaperone has discrete domains required for SseB stabilization and export, and binds within the C-terminus of SseB and SseD Free

Abstract

SseA, a key virulence determinant, is a small, basic pI protein encoded within the pathogenicity island 2 and serves as a type III secretion system chaperone for SseB and SseD. Both SseA partners are subunits of the surface-localized translocon module that delivers effectors into the host cell; SseB is predicted to compose the translocon sheath and SseD is a putative translocon pore subunit. In this study, SseA molecular interactions with its partners were characterized further. Yeast two-hybrid screens indicate that SseA binding requires a C-terminal domain within both partners. An additional central domain within SseD was found to influence binding. The SseA-binding region within SseB was found to encompass a predicted amphipathic helix of a type participating in coiled-coil interactions that are implicated in the assembly of translocon sheaths. Deletions that impinge upon this putative coiled-coiled domain prevent SseA binding, suggesting that SseA occupies a portion of the coiled-coil. SseA occupancy of this motif is envisioned to be sufficient to prevent premature SseB self-association inside bacteria. Domain mapping on the chaperone was also performed. A deletion of the SseA N-terminus, or site-directed mutations within this region, allowed stabilization of SseB, but its export was disrupted. Therefore, the N-terminus of SseA provides a function that is essential for SseB export, but dispensable for partner binding and stabilization.

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  • Published Online:
Keyword(s): EPEC, enteropathogenic E. coli , GST, glutathione S-transferase , LEE, locus for enterocyte effacement , MCS, multiple cloning site , SPI2, Salmonella pathogenicity island 2 and TTS, type III secretion
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2004-07-01
2024-04-25
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The SPI2-encoded SseA chaperone has discrete domains required for SseB stabilization and export, and binds within the C-terminus of SseB and SseD
Microbiology 150, 2055 (2004); https://doi.org/10.1099/mic.0.26997-0
/content/journal/micro/10.1099/mic.0.26997-0
/content/journal/micro/10.1099/mic.0.26997-0
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