Back to Search
Start Over
Resolution of Holliday junctions in vitro requires the Escherichia coli ruvC gene product.
- Source :
-
Proceedings of the National Academy of Sciences of the United States of America [Proc Natl Acad Sci U S A] 1991 Jul 15; Vol. 88 (14), pp. 6063-7. - Publication Year :
- 1991
-
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.
- Subjects :
- Bacteriophage phi X 174 genetics
Base Sequence
DNA, Viral genetics
Gene Expression
Molecular Sequence Data
Oligonucleotide Probes
Plasmids
Bacterial Proteins genetics
DNA Replication
DNA, Bacterial genetics
Endodeoxyribonucleases
Escherichia coli genetics
Escherichia coli Proteins
Genes, Bacterial
Subjects
Details
- Language :
- English
- ISSN :
- 0027-8424
- Volume :
- 88
- Issue :
- 14
- Database :
- MEDLINE
- Journal :
- Proceedings of the National Academy of Sciences of the United States of America
- Publication Type :
- Academic Journal
- Accession number :
- 1829835
- Full Text :
- https://doi.org/10.1073/pnas.88.14.6063