title: Disruption of IcsP, the major Shigella protease that cleaves IcsA, accelerates actin-based motility.
abstract: Shigella pathogenesis involves bacterial invasion of colonic epithelial cells and movement of bacteria through the cytoplasm and into adjacent cells by means of actin-based motility. The Shigella protein IcsA (VirG) is unipolar on the bacterial surface and is both necessary and sufficient for actin-based motility. IcsA is inserted into the outer membrane as a 120-kDa polypeptide that is subsequently slowly cleaved, thereby releasing the 95-kDa amino-terminal portion into the culture supernatant. IcsP, the major Shigella protease that cleaves IcsA, was identified and cloned. It has significant sequence similarity to the E. coli serine proteases, OmpP and OmpT. Disruption of icsP in serotype 2a S. flexneri leads to a marked reduction in IcsA cleavage, increased amounts of IcsA associated with the bacterium and altered distribution of IcsA on the bacterial surface. The icsP mutant displays significantly increased rates of actin-based motility, with a mean speed 27% faster than the wild-type strain; moreover, a significantly greater percentage of the icsP mutant moves in the cytoplasm. Yet, plaque formation on epithelial monolayers by the mutant was not altered detectably. These data suggest that IcsA, and not a host protein, is limiting in the rate of actin-based motility of wild-type serotype 2a S. flexneri.
authors.affiliation: Department of Microbiology and Immunology, Albert Einstein College of Medicine, Bronx, NY 10461-1602, USA.
authors.forename: K D
authors.initials: KD
authors.lastname: Shere
authors.forename: S
authors.initials: S
authors.lastname: Sallustio
authors.forename: A
authors.initials: A
authors.lastname: Manessis
authors.forename: T G
authors.initials: TG
authors.lastname: D'Aversa
authors.forename: M B
authors.initials: MB
authors.lastname: Goldberg
mesh_terms: D000199:Actins / Q000378:metabolism*; D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism*; D003001:Cloning, Molecular; D004268:DNA-Binding Proteins / Q000378:metabolism*; D005798:Genes, Bacterial; D008969:Molecular Sequence Data; D009068:Movement / Q000502:physiology; D009154:Mutation; D017386:Sequence Homology, Amino Acid; D012763:Shigella flexneri / Q000235:genetics / Q000472:pathogenicity / Q000502:physiology*; D014157:Transcription Factors / Q000378:metabolism*
chemical_list: D000199:Actins; D001426:Bacterial Proteins; D004268:DNA-Binding Proteins; D014157:Transcription Factors; C058483:virG protein, Shigella flexneri; C060971:SopA protein, Bacteria