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

TY - JOUR

T1 - A RecB-family nuclease motif in the Type I restriction endonuclease EcoR124I

AU - Šišǎkovǎ, Eva

AU - Stanley, Louise K.

AU - Weiserovǎ, Marie

AU - Szczelkun, Mark D.

N1 - Funding Information: We thank Mark Dillingham, Frank Peske and Ralf Seidel for comments on the article, and Mark Sanderson for preliminary experiments on EcoAI. This work was supported by a Wellcome Trust Senior Research Fellowship in Basic Biomedical Sciences (067439 to M.D.S.); The Grant Agency of the Czech Republic (204/07/0325 to M.W.); and, The European Commission through the BioNano-Switch project (043288 to M.W. and E.S.). Funding to pay the Open Access publication charges for this article was provided by Wellcome Trust Value-in-People Award 078595.

PY - 2008/7/1

Y1 - 2008/7/1

N2 - 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.

AB - 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.

UR - https://www.scopus.com/pages/publications/47249088354

U2 - 10.1093/nar/gkn333

DO - 10.1093/nar/gkn333

M3 - Article

C2 - 18511464

AN - SCOPUS:47249088354

SN - 0305-1048

VL - 36

SP - 3939

EP - 3949

JO - Nucleic Acids Research

JF - Nucleic Acids Research

IS - 12

ER -