A RecB-family nuclease motif in the Type I restriction endonuclease EcoR124I

Eva Šišǎkovǎ, Louise K. Stanley, Marie Weiserovǎ, Mark D. Szczelkun (Lead / Corresponding author)

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    Abstract

    The Type I restriction-modification enzyme EcoR124I is an ATP-dependent endonuclease that uses dsDNA translocation to locate and cleave distant non-specific DNA sites. Bioinformatic analysis of the HsdR subunits of EcoR124I and related Type I enzymes showed that in addition to the principal PD-(E/D)xK Motifs, I, II and III, a QxxxY motif is also present that is characteristic of RecB-family nucleases. The QxxxY motif resides immediately C-terminal to Motif III within a region of predicted α-helix. Using mutagenesis, we examined the role of the Q and Y residues in DNA binding, translocation and cleavage. Roles for the QxxxY motif in coordinating the catalytic residues or in stabilizing the nuclease domain on the DNA are discussed.

    Original languageEnglish
    Pages (from-to)3939-3949
    Number of pages11
    JournalNucleic Acids Research
    Volume36
    Issue number12
    Early online date29 May 2008
    DOIs
    Publication statusPublished - 1 Jul 2008

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