Please use this identifier to cite or link to this item:
https://hdl.handle.net/2440/79619
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Type: | Journal article |
Title: | Identification of Shigella flexneri IcsA residues affecting interaction with N-WASP, and evidence for IcsA-IcsA co-operative interaction |
Author: | Teh, M. Morona, R. |
Citation: | PLoS One, 2013; 8(2):1-17 |
Publisher: | Public Library of Science |
Issue Date: | 2013 |
ISSN: | 1932-6203 1932-6203 |
Editor: | Adler, B. |
Abstract: | The Shigella flexneri IcsA (VirG) protein is a polarly distributed outer membrane protein that is a fundamental virulence factor which interacts with neural Wiskott-Aldrich syndrome protein (N-WASP). The activated N-WASP then activates the Arp2/3 complex which initiates de novo actin nucleation and polymerisation to form F-actin comet tails and allows bacterial cell-to-cell spreading. In a previous study, IcsA was found to have three N-WASP interacting regions (IRs): IR I (aa 185–312), IR II (aa 330–382) and IR III (aa 508–730). The aim of this study was to more clearly define N-WASP interacting regions II and III by site-directed mutagenesis of specific amino acids. Mutant IcsA proteins were expressed in both smooth lipopolysaccharide (S-LPS) and rough LPS (R-LPS) S. flexneri strains and characterised for IcsA production level, N-WASP recruitment and F-actin comet tail formation. We have successfully identified new amino acids involved in N-WASP recruitment within different N-WASP interacting regions, and report for the first time using co-expression of mutant IcsA proteins, that N-WASP activation involves interactions with different regions on different IcsA molecules as shown by Arp3 recruitment. In addition, our findings suggest that autochaperone (AC) mutant protein production was not rescued by another AC region provided in trans, differing to that reported for two other autotransporters, PrtS and BrkA autotransporters. |
Keywords: | Shigella flexneri Actins Lipopolysaccharides Bacterial Proteins DNA-Binding Proteins Transcription Factors Mutagenesis, Site-Directed Mutation Wiskott-Aldrich Syndrome Protein, Neuronal Actin-Related Protein 3 Protein Interaction Domains and Motifs |
Rights: | Copyright: © 2013 Teh, Morona. 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 author and source are credited. |
DOI: | 10.1371/journal.pone.0055152 |
Published version: | http://dx.doi.org/10.1371/journal.pone.0055152 |
Appears in Collections: | Aurora harvest Molecular and Biomedical Science publications |
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hdl_79619.pdf | Published version | 4.14 MB | Adobe PDF | View/Open |
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