Potential double-flipping mechanism by E. coli MutY

Paul G. House, David E. Volk, Varatharasa Thiviyanathan, Raymond C. Manuel, Bruce A. Luxon, David G. Gorenstein, R. Stephen Lloyd

    Research output: Contribution to journalArticle

    12 Citations (Scopus)

    Abstract

    To understand the structural basis of the recognition and removal of specific mismatched bases in double-stranded DNAs by the DNA repair glycosylase MutY, a series of structural and functional analyses have been conducted. MutY is a 39-kDa enzyme from Escherichia coli, which to date has been refractory to structural determination in its native, intact conformation. However, following limited proteolytic digestion, it was revealed that the MutY protein is composed of two modules, a 26-kDa domain that retains essential catalytic function (designated p26MutY) and a 13-kDa domain that is implicated in substrate specificity and catalytic efficiency. Several structures of the 26-kDa domain have been solved by X-ray crystallographic methods to a resolution of up to 1.2 Å. The structure of a catalytically incompetent mutant of p26MutY complexed with an adenine in the substrate-binding pocket allowed us to propose a catalytic mechanism for MutY. Since reporting the structure of p26MutY, significant progress has been made in solving the solution structure of the noncatalytic C-terminal 13-kDa domain of MutY by NMR spectroscopy. The topology and secondary structure of this domain are very similar to that of MutT, a pyrophosphohydrolase. Molecular modeling techniques employed to integrate the two domains of MutY with DNA suggest that MutY can wrap around the DNA and initiate catalysis by potentially flipping adenine and 8-oxoguanine out of the DNA helix.

    Original languageEnglish (US)
    Pages (from-to)349-364
    Number of pages16
    JournalProgress in Nucleic Acid Research and Molecular Biology
    Volume68
    StatePublished - 2001

    Fingerprint

    Escherichia coli
    DNA
    Adenine
    DNA Glycosylases
    Substrate Specificity
    Catalysis
    DNA Repair
    Digestion
    Magnetic Resonance Spectroscopy
    X-Rays
    Enzymes
    Proteins
    8-hydroxyguanine

    ASJC Scopus subject areas

    • Molecular Biology

    Cite this

    House, P. G., Volk, D. E., Thiviyanathan, V., Manuel, R. C., Luxon, B. A., Gorenstein, D. G., & Stephen Lloyd, R. (2001). Potential double-flipping mechanism by E. coli MutY. Progress in Nucleic Acid Research and Molecular Biology, 68, 349-364.

    Potential double-flipping mechanism by E. coli MutY. / House, Paul G.; Volk, David E.; Thiviyanathan, Varatharasa; Manuel, Raymond C.; Luxon, Bruce A.; Gorenstein, David G.; Stephen Lloyd, R.

    In: Progress in Nucleic Acid Research and Molecular Biology, Vol. 68, 2001, p. 349-364.

    Research output: Contribution to journalArticle

    House, PG, Volk, DE, Thiviyanathan, V, Manuel, RC, Luxon, BA, Gorenstein, DG & Stephen Lloyd, R 2001, 'Potential double-flipping mechanism by E. coli MutY', Progress in Nucleic Acid Research and Molecular Biology, vol. 68, pp. 349-364.
    House PG, Volk DE, Thiviyanathan V, Manuel RC, Luxon BA, Gorenstein DG et al. Potential double-flipping mechanism by E. coli MutY. Progress in Nucleic Acid Research and Molecular Biology. 2001;68:349-364.
    House, Paul G. ; Volk, David E. ; Thiviyanathan, Varatharasa ; Manuel, Raymond C. ; Luxon, Bruce A. ; Gorenstein, David G. ; Stephen Lloyd, R. / Potential double-flipping mechanism by E. coli MutY. In: Progress in Nucleic Acid Research and Molecular Biology. 2001 ; Vol. 68. pp. 349-364.
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