Abstract
Asymmetric localization is key to the proper function of certain prokaryotic proteins important to virulence, chemotaxis, cell division, development, motility, and adhesion. Shigella IcsA is localized to the old pole of the bacterium, where it mediates assembly of an actin tail inside infected mammalian cells. IcsA (VirG) is essential to Shigella intracellular motility and virulence. We used translational fusions between portions of IcsA and the green fluorescent protein (GFP) to determine the regions of IcsA that are necessary and sufficient for its targeting to the bacterial old pole. An IcsA-GFP fusion that lacks a signal peptide localized to the old pole, indicating that signal peptide-mediated secretion is not required for polar localization. Two regions within IcsA were required for localization of an IcsA-GFP fusion to the old pole. Further characterization of these regions indicated that amino acids 1-104 and 507-620 were each independently sufficient for polar localization. Finally, when expressed in Escherichia coli, Salmonella typhimurium, Yersinia pseudotuberculosis, and Vibrio cholerae, each of the two targeting regions localized to the pole, indicating that the mechanism of polar targeting used by IcsA is present generally among Enterobacteriacae and Vibrio.
MeSH Terms
Amino Acid Sequence
Bacterial Proteins/chemistry,genetics,metabolism
Cell Polarity
DNA-Binding Proteins/chemistry,genetics,metabolism
Enterobacteriaceae/genetics,metabolism,ultrastructure
Escherichia coli/genetics,metabolism,ultrastructure
Genes, Reporter
Green Fluorescent Proteins
Luminescent Proteins/analysis,metabolism
Molecular Sequence Data
Protein Structure, Tertiary
Protein Transport
Recombinant Fusion Proteins/metabolism
Salmonella typhimurium/genetics,metabolism,ultrastructure
Shigella flexneri/genetics,metabolism,pathogenicity,ultrastructure
Species Specificity
Transcription Factors/chemistry,genetics,metabolism
Vibrio cholerae/genetics,metabolism,ultrastructure
Virulence/genetics
Yersinia pseudotuberculosis/genetics,metabolism,ultrastructure
Chemicals
Bacterial Proteins
DNA-Binding Proteins
Luminescent Proteins
Recombinant Fusion Proteins
Transcription Factors
virG protein, Shigella flexneri
Green Fluorescent Proteins
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Charles M
Department of Microbiology and Immunology, Albert Einstein College of Medicine, Bronx, NY 10461, USA.
Pérez M
Kobil J H
Goldberg M B
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