Resolution of a Holliday junction by vaccinia topoisomerase requires a spacer DNA segment 3' of the CCCTT↓ cleavage sites Journal Article
| Authors: | Sekiguchi, J.; Cheng, C.; Shuman, S. |
| Article Title: | Resolution of a Holliday junction by vaccinia topoisomerase requires a spacer DNA segment 3' of the CCCTT↓ cleavage sites |
| Abstract: | Vaccinia virus DNA topoisomerase catalyzes resolution of synthetic Holliday junctions in vitro. The mechanism entails concerted transesterifications at two recognition sites, 5'-CCCTT↓, that are opposed within a partially mobile four-way junction. Efficient resolution occurs on a junction with a 10 bp segment of branch mobility (5'-GCCCTTATCG) that extends 4 bp 3' of the scissile phosphate. Here we report that resolution is decreased when branch mobility is limited to an 8 bp segment extending 2 bp 3' of the cleavage site and then eliminated when branch mobility is confined to the 6 bp GCCCTT sequence 5' of the scissile phosphate. We surmise that a spacer region 3' of CCCTT is needed for simultaneous cleavage at two opposing sites at the junction. Branch mobility is bot required for reaction chemistry at a junction, becausetopoisomerase cleaves a single CCCTT site in a non-mobile four-way junction where the scissile phosphate is at the cross-over point. The junction resolvase activity of topoisomerase may be involved in forming the hairpin telomeres of the vaccinia genome. |
| Keywords: | controlled study; nonhuman; dna recombination; telomere; protein binding; in vitro study; dose-response relationship, drug; animalia; dna; molecular sequence data; dna viruses; vaccinia virus; base sequence; binding site; dna sequence; binding sites; esterification; catalysis; nucleic acid conformation; phosphate; dna cleavage; dna topoisomerase; dna topoisomerases, type i; vaccinia; virus gene; holliday junction; spacer dna; priority journal; article; resolvase; dna transfer |
| Journal Title: | Nucleic Acids Research |
| Volume: | 28 |
| Issue: | 14 |
| ISSN: | 0305-1048 |
| Publisher: | Oxford University Press |
| Date Published: | 2000-07-15 |
| Start Page: | 2658 |
| End Page: | 2663 |
| Language: | English |
| PUBMED: | 10908320 |
| PROVIDER: | scopus |
| PMCID: | PMC102645 |
| DOI: | 10.1093/nar/28.14.2658 |
| DOI/URL: | |
| Notes: | Export Date: 18 November 2015 -- Source: Scopus |
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