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Article ; Online: Interaction of branch migration translocases with the Holliday junction-resolving enzyme and their implications in Holliday junction resolution.

Cañas, Cristina / Suzuki, Yuki / Marchisone, Chiara / Carrasco, Begoña / Freire-Benéitez, Verónica / Takeyasu, Kunio / Alonso, Juan C / Ayora, Silvia

The Journal of biological chemistry

2014  Volume 289, Issue 25, Page(s) 17634–17646

Abstract: ... suggested that the absence of the RuvAB branch migration translocase and the RecU HJ resolvase is ... of HJ branch migration and/or resolution are poorly characterized in Firmicutes. Genetic evidence ... a ternary complex. RecU cannot stimulate the ATPase or branch migration activity of RuvB. The presence ...

Abstract Double-strand break repair involves the formation of Holliday junction (HJ) structures that need to be resolved to promote correct replication and chromosomal segregation. The molecular mechanisms of HJ branch migration and/or resolution are poorly characterized in Firmicutes. Genetic evidence suggested that the absence of the RuvAB branch migration translocase and the RecU HJ resolvase is synthetically lethal in Bacillus subtilis, whereas a recU recG mutant was viable. In vitro RecU, which is restricted to bacteria of the Firmicutes phylum, binds HJs with high affinity. In this work we found that RecU does not bind simultaneously with RecG to a HJ. RuvB by interacting with RecU bound to the central region of HJ DNA, loses its nonspecific association with DNA, and re-localizes with RecU to form a ternary complex. RecU cannot stimulate the ATPase or branch migration activity of RuvB. The presence of RuvB·ATPγS greatly stimulates RecU-mediated HJ resolution, but the addition of ATP or RuvA abolishes this stimulatory effect. A RecU·HJ·RuvAB complex might be formed. RecU does not increase the RuvAB activities but slightly inhibits them.
MeSH term(s) Bacillus subtilis/enzymology ; Bacillus subtilis/genetics ; Bacterial Proteins/genetics ; Bacterial Proteins/metabolism ; DNA, Bacterial/genetics ; DNA, Bacterial/metabolism ; DNA, Cruciform/genetics ; DNA, Cruciform/metabolism ; Exodeoxyribonucleases/genetics ; Exodeoxyribonucleases/metabolism
Chemical Substances Bacterial Proteins ; DNA, Bacterial ; DNA, Cruciform ; RuvB protein, Bacteria ; Exodeoxyribonucleases (EC 3.1.-) ; recJ protein, Bacteria (EC 3.1.11.-)
Language English
Publishing date 2014-04-25
Publishing country United States
Document type Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID 2997-x
ISSN 1083-351X ; 0021-9258
ISSN (online) 1083-351X
ISSN 0021-9258
DOI 10.1074/jbc.M114.552794
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