Home LiteratureArticle Details
PMID: 11481451 Published · ppublish English Comparative Study Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Polar targeting of Shigella virulence factor IcsA in Enterobacteriacae and Vibrio.

Charles M, Pérez M, Kobil JH, Goldberg MB

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
References (34)
34 references, click to expand
  1. Identification of icsA, a plasmid locus of Shigella flexneri that governs bacterial intra- and intercellular spread through interaction with F-actin.
    Proc Natl Acad Sci U S A. 1989 May;86(10):3867-71 PMID: 2542950
  2. Polar location of the chemoreceptor complex in the Escherichia coli cell.
    Science. 1993 Mar 19;259(5102):1717-23 PMID: 8456299
  3. MinDE-dependent pole-to-pole oscillation of division inhibitor MinC in Escherichia coli.
    J Bacteriol. 1999 Oct;181(20):6419-24 PMID: 10515933
  4. Type IV pili and cell motility.
    Mol Microbiol. 1999 Apr;32(1):1-10 PMID: 10216854
  5. Establishment of unipolar localization of IcsA in Shigella flexneri 2a is not dependent on virulence plasmid determinants.
    Infect Immun. 1999 Jan;67(1):350-6 PMID: 9864236
  6. Polar localization of a bacterial chemoreceptor.
    Genes Dev. 1992 May;6(5):825-36 PMID: 1577276
  7. A set of compatible tac promoter expression vectors.
    Gene. 1996 Oct 24;177(1-2):133-6 PMID: 8921858
  8. Periplasmic transit and disulfide bond formation of the autotransported Shigella protein IcsA.
    J Bacteriol. 2001 Feb;183(3):951-8 PMID: 11208794
  9. The unipolar Shigella surface protein IcsA is targeted directly to the bacterial old pole: IcsP cleavage of IcsA occurs over the entire bacterial surface.
    Mol Microbiol. 1999 Apr;32(2):367-77 PMID: 10231492
  10. Vaccination against shigellosis with attenuated Shigella flexneri 2a strain SC602.
    Infect Immun. 1999 Jul;67(7):3437-43 PMID: 10377124
  11. Functional analysis of a rickettsial OmpA homology domain of Shigella flexneri icsA.
    J Bacteriol. 1999 Feb;181(3):869-78 PMID: 9922250
  12. Differential localization of two histidine kinases controlling bacterial cell differentiation.
    Mol Cell. 1999 Nov;4(5):683-94 PMID: 10619016
  13. Cholera toxin genes: nucleotide sequence, deletion analysis and vaccine development.
    Nature. 1983 Dec 8-14;306(5943):551-7 PMID: 6646234
  14. Unipolar localization and ATPase activity of IcsA, a Shigella flexneri protein involved in intracellular movement.
    J Bacteriol. 1993 Apr;175(8):2189-96 PMID: 8468279
  15. Functional analysis of Shigella VirG domains essential for interaction with vinculin and actin-based motility.
    J Biol Chem. 1996 Sep 6;271(36):21878-85 PMID: 8702989
  16. Construction and properties of a family of pACYC184-derived cloning vectors compatible with pBR322 and its derivatives.
    Gene. 1991 Jun 15;102(1):75-8 PMID: 1840539
  17. A novel family of channel-forming, autotransporting, bacterial virulence factors.
    Mol Membr Biol. 1997 Jul-Sep;14(3):113-23 PMID: 9394291
  18. virG, a plasmid-coded virulence gene of Shigella flexneri: identification of the virG protein and determination of the complete coding sequence.
    J Bacteriol. 1989 Jan;171(1):353-9 PMID: 2644195
  19. FACS-optimized mutants of the green fluorescent protein (GFP).
    Gene. 1996;173(1 Spec No):33-8 PMID: 8707053
  20. Extracellular transport of VirG protein in Shigella.
    J Biol Chem. 1995 Dec 29;270(52):30874-80 PMID: 8537341
  21. SopA, the outer membrane protease responsible for polar localization of IcsA in Shigella flexneri.
    Mol Microbiol. 1997 Mar;23(5):1063-73 PMID: 9076742
  22. Secretion of beta-lactamase requires the carboxy end of the protein.
    Cell. 1980 Jul;20(3):749-60 PMID: 6448092
  23. DIALIGN 2: improvement of the segment-to-segment approach to multiple sequence alignment.
    Bioinformatics. 1999 Mar;15(3):211-8 PMID: 10222408
  24. Tight regulation, modulation, and high-level expression by vectors containing the arabinose PBAD promoter.
    J Bacteriol. 1995 Jul;177(14):4121-30 PMID: 7608087
  25. OmpB (osmo-regulation) and icsA (cell-to-cell spread) mutants of Shigella flexneri: vaccine candidates and probes to study the pathogenesis of shigellosis.
    Vaccine. 1991 Jun;9(6):416-22 PMID: 1887672
  26. EPITHELIAL CELL PENETRATION AS AN ESSENTIAL STEP IN THE PATHOGENESIS OF BACILLARY DYSENTERY.
    J Bacteriol. 1964 Nov;88(5):1503-18 PMID: 16562000
  27. Topological regulation of cell division in Escherichia coli involves rapid pole to pole oscillation of the division inhibitor MinC under the control of MinD and MinE.
    Mol Microbiol. 1999 Oct;34(1):82-90 PMID: 10540287
  28. Presence of a virulence-associated plasmid in Yersinia pseudotuberculosis.
    Infect Immun. 1980 Jun;28(3):1044-7 PMID: 6249747
  29. Membrane redistribution of the Escherichia coli MinD protein induced by MinE.
    J Bacteriol. 2000 Feb;182(3):613-9 PMID: 10633093
  30. Cell cycle-dependent polar localization of an essential bacterial histidine kinase that controls DNA replication and cell division.
    Cell. 1999 Apr 2;97(1):111-20 PMID: 10199407
  31. A genetic determinant required for continuous reinfection of adjacent cells on large plasmid in S. flexneri 2a.
    Cell. 1986 Aug 15;46(4):551-5 PMID: 3524856
  32. Disruption of IcsP, the major Shigella protease that cleaves IcsA, accelerates actin-based motility.
    Mol Microbiol. 1997 Aug;25(3):451-62 PMID: 9302008
  33. The great escape: structure and function of the autotransporter proteins.
    Trends Microbiol. 1998 Sep;6(9):370-8 PMID: 9778731
  34. Rapid pole-to-pole oscillation of a protein required for directing division to the middle of Escherichia coli.
    Proc Natl Acad Sci U S A. 1999 Apr 27;96(9):4971-6 PMID: 10220403
Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
0027-8424
Published
2001-08-14
Epub
2001-00-31
Pages
9871-6
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC55545
Subset
IM
Grants
NIGMS NIH HHS · GM16654 · United States
NIAID NIH HHS · AI35817 · United States
NHLBI NIH HHS · T32 HL007118 · United States
NIGMS NIH HHS · F31 GM016654 · United States
NHLBI NIH HHS · HL07118 · United States
NIAID NIH HHS · R01 AI035817 · United States
Analysis Services
Analysis Services

Contact

No. 2 Wenbo Road, Zhangqiu District, Jinan, Shandong

Qilu Normal University · Genelibs Bioinformatics Lab

750 Shunhua Rd, Jinan

2F, Bldg F, University Science Park

Tel: 0531-88819269

WeChat Official Account

Follow our WeChat subscription account for real-time updates and the latest in medical and biological research.


Business Email

E-mail: product@genelibs.com