Resolution of Holliday junctions in vitro requires the Escherichia coli ruvC gene product.
- 15 July 1991
- journal article
- Published by Proceedings of the National Academy of Sciences in Proceedings of the National Academy of Sciences
- Vol. 88 (14), 6063-6067
- https://doi.org/10.1073/pnas.88.14.6063
Abstract
In previous studies, Holliday junctions generated during RecA-mediated strand-exchange reactions were resolved by fractionated Escherichia coli extracts. We now report the specific binding and cleavage of synthetic Holliday junctions (50 base pairs long) by a fraction purified by chromatography on DEAE-cellulose, phosphocellulose, and single-stranded DNA-cellulose. The cleavage reaction provided a sensitive assay with which to screen extracts prepared from recombination/repair-deficient mutants. Cells with mutations in ruvC lack the nuclease activity that cleaves synthetic Holliday junctions in vitro. This deficiency was restored by a multicopy plasmid carrying a ruvC+ gene that overexpressed junction-resolving activity. The UV sensitivity and deficiency in recombinational repair of DNA exhibited by ruv mutants lead us to suggest that RuvC resolves Holliday junctions in vivo.Keywords
This publication has 25 references indexed in Scilit:
- Evidence of abortive recombination in ruv mutants of Escherichia coli K12Molecular Genetics and Genomics, 1991
- Purification and preliminary characterization of the Escherichia coli K-12 recF proteinJournal of Bacteriology, 1990
- Overproduction, purification, and ATPase activity of the Escherichia coli RuvB protein involved in DNA repairJournal of Bacteriology, 1989
- Identification and purification of a single-stranded-DNA-specific exonuclease encoded by the recJ gene of Escherichia coli.Proceedings of the National Academy of Sciences, 1989
- Identification of the recR locus of Escherichia coli K-12 and analysis of its role in recombination and DNA repairMolecular Genetics and Genomics, 1989
- Structure and regulation of the Escherichia coli ruv operon involved in DNA repair and recombinationJournal of Bacteriology, 1988
- Nucleotide sequencing of theruvregion ofEscherichia coliK-12 reveals a LexA regulated operon encoding two genesNucleic Acids Research, 1988
- Genetic Analysis of Conjugational Recombination in Escherichia coli K12 Strains Deficient in RecBCD EnzymeMicrobiology, 1987
- ENZYMES OF GENERAL RECOMBINATIONAnnual Review of Biochemistry, 1987
- Chi-stimulated patches are heteroduplex, with recombinant information on the phage λ r chainCell, 1987