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Within host cells, IcsA activates the host actin regulatory protein, neural Wiskott-Aldrich syndrome protein (N-WASP), which in turn recruits the Arp2\/3 complex, which nucleates host actin to form F-actin comet tails and initiate bacterial motility. Linker insertion mutagenesis was undertaken to randomly introduce 5-amino-acid in-frame insertions within IcsA. Forty-seven linker insertion mutants were isolated and expressed in<jats:italic>S. flexneri<\/jats:italic>\u0394<jats:italic>icsA<\/jats:italic>strains. Mutants were characterized for IcsA protein production, cell surface expression and localization, intercellular spreading, F-actin comet tail formation, and N-WASP recruitment. Using this approach, we have identified a putative autochaperone region required for IcsA biogenesis, and our data suggest an additional region, not previously identified, is required for N-WASP recruitment.<\/jats:p>","DOI":"10.1128\/jb.00093-08","type":"journal-article","created":{"date-parts":[[2008,5,4]],"date-time":"2008-05-04T01:46:16Z","timestamp":1209865576000},"page":"4666-4676","update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":42,"title":["Mutagenesis of the<i>Shigella flexneri<\/i>Autotransporter IcsA Reveals Novel Functional Regions Involved in IcsA Biogenesis and Recruitment of Host Neural Wiscott-Aldrich Syndrome Protein"],"prefix":"10.1128","volume":"190","author":[{"given":"Kerrie L.","family":"May","sequence":"first","affiliation":[{"name":"Australian Bacterial Pathogenesis Program, Discipline of Microbiology and Immunology, School of Molecular and Biomedical Science, University of Adelaide, South Australia, Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Renato","family":"Morona","sequence":"additional","affiliation":[{"name":"Australian Bacterial Pathogenesis Program, Discipline of Microbiology and Immunology, School of Molecular and Biomedical Science, University of Adelaide, South Australia, Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"235","reference":[{"key":"e_1_3_2_2_2","doi-asserted-by":"crossref","first-page":"367","DOI":"10.1099\/13500872-145-2-367","volume":"145","year":"1999","unstructured":"Baker, S., J. 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