The identification of exported proteins with gene fusions to invasin

Micah J. Worley, Igor Stojiljkovic, Fred Heffron

    Research output: Contribution to journalArticle

    8 Citations (Scopus)

    Abstract

    Exported proteins are integral to understanding the biology of bacterial organisms. They have special significance in pathogenesis research because they can mediate critical interactions between pathogens and eukaryotic cell surfaces. Further, they frequently serve as targets for vaccines and diagnostic tests. The commonly used genetic assays for identifying exported proteins use fusions to alkaline phosphatase or beta-lactamase. These systems are not ideal for identifying outer membrane proteins because they identify a large number of inner membrane proteins as well. We addressed this problem by developing a gene fusion system that preferentially identifies proteins that contain cleavable signal sequences and are released from the inner membrane. This system selects fusions that restore outer membrane localization to an amino terminal-truncated Yersinia pseudotuberculosis invasin derivative. In the present study, a variety of Salmonella typhimurium proteins that localize beyond the inner membrane were identified with gene fusions to this invasin derivative. Previously undescribed proteins identified include ones that share homology with components of fimbrial operons, multiple drug resistance efflux pumps and a haemolysin. All of the positive clones analysed contain cleavable signal sequences. Moreover, over 40% of the genes identified encode putative outer membrane proteins. This system has several features that may make it especially useful in the study of genetically intractable organisms.

    Original languageEnglish (US)
    Pages (from-to)1471-1480
    Number of pages10
    JournalMolecular Microbiology
    Volume29
    Issue number6
    DOIs
    StatePublished - 1998

    Fingerprint

    Gene Fusion
    Membrane Proteins
    Protein Sorting Signals
    Proteins
    Membranes
    Yersinia pseudotuberculosis
    Hemolysin Proteins
    Multiple Drug Resistance
    Eukaryotic Cells
    Salmonella typhimurium
    beta-Lactamases
    Operon
    Routine Diagnostic Tests
    Alkaline Phosphatase
    Vaccines
    Clone Cells
    Research
    Genes

    ASJC Scopus subject areas

    • Molecular Biology
    • Microbiology

    Cite this

    The identification of exported proteins with gene fusions to invasin. / Worley, Micah J.; Stojiljkovic, Igor; Heffron, Fred.

    In: Molecular Microbiology, Vol. 29, No. 6, 1998, p. 1471-1480.

    Research output: Contribution to journalArticle

    Worley, Micah J. ; Stojiljkovic, Igor ; Heffron, Fred. / The identification of exported proteins with gene fusions to invasin. In: Molecular Microbiology. 1998 ; Vol. 29, No. 6. pp. 1471-1480.
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