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Monitoring the Assembly of a Secreted Bacterial Virulence Factor Using Site-specific Crosslinking
Published on: December 17, 2013
Assembly of pili on the surface of Bacillus cereus vegetative cells
Jonathan M Budzik1, Luciano A Marraffini, Olaf Schneewind
1Department of Microbiology, University of Chicago, Chicago, IL 60637, USA.
Abstract:
Vegetative forms of Bacillus cereus are reported to form pili, thin protein filaments that protrude up to 1 mum from the bacterial surface. Pili are assembled from two precursor proteins, BcpA and BcpB, in a manner requiring a pilus-associated sortase enzyme (SrtD). Pili are also formed on the surface of Bacillus anthracis expressing bcpA-srtD-bcpB. BcpA is distributed throughout the entire pilus, whereas BcpB appears positioned at its tip. In agreement with the hypothesis for pilus assembly in Gram-positive bacteria, BcpA encompasses the YPK pilin motif and the LPXTG sorting signal, each of which is absolutely required for the incorporation of BcpA and BcpB into pili. In contrast to BcpB, which relies on the presence of BcpA for incorporation into pili, BcpA fibre assembly occurs even in the absence of BcpB. B. anthracis sortase A (srtA), but not sortase B (srtB) or C (srtC), is required for proper anchoring of pili to the bacterial envelope, suggesting that BcpA/BcpB pili are linked to peptidoglycan cross-bridges.
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