PMID:8393934
| Citation |
Parsons, CA and West, SC (1993) Formation of a RuvAB-Holliday junction complex in vitro. J. Mol. Biol. 232:397-405 |
|---|---|
| Abstract |
The ruvA and ruvB genes of Escherichia coli encode a novel DNA helicase that interacts with Holliday junctions and promotes branch migration. In this work, we have investigated the protein-DNA complexes formed between RuvA, RuvB and Holliday junctions. As shown previously, RuvA protein binds a synthetic Holliday junction in vitro, to form a specific protein-DNA complex that can be detected by a band-shift assay. We now show that the combined presence of RuvA and RuvB results in a super-shift of this complex indicative of the formation of a RuvAB-Holliday junction complex. In the absence of RuvA, the RuvB protein fails to bind Holliday junctions. The RuvAB-Holliday junction complex was detected by the band-shift assay only under conditions that favoured its stability, e.g. complex formation in the presence of a nucleoside triphosphate that can not be hydrolysed by RuvB (adenosine 5'-[gamma-thio]triphosphate). In contrast, nucleoside triphosphates that can be hydrolysed (ATP, dATP, dCTP or TTP), lead to RuvAB-mediated branch migration of the junction. These results indicate that the formation of a (RuvAB-ATP)-Holliday junction complex represents the first step in the process of branch migration, and that branch migration is dependent upon ATP hydrolysis. In addition, we show that Holliday junction DNA stimulates the ATPase activity of RuvAB to a greater extent than either single-stranded or linear duplex DNA. |
| Links |
PubMed Online version:10.1006/jmbi.1993.1399 |
| Keywords |
Adenosine Triphosphatases/analysis; Adenosine Triphosphate/analogs & derivatives; Adenosine Triphosphate/metabolism; Bacterial Proteins/metabolism; Base Sequence; DNA Helicases/metabolism; DNA Repair; DNA, Bacterial/genetics; DNA, Bacterial/metabolism; DNA-Binding Proteins/metabolism; Deoxyguanine Nucleotides/metabolism; Escherichia coli/genetics; Escherichia coli/metabolism; Escherichia coli Proteins; Models, Genetic; Molecular Sequence Data; Recombination, Genetic |
| edit table |
Significance
You can help EcoliWiki by summarizing why this paper is useful
Useful Materials and Methods
You can help Ecoliwiki by describing the useful materials (strains, plasmids, antibodies, etc) described in this paper.
Annotations
<annotationlinks/>
EcoliWiki Links
Add links to pages that link here (e.g. gene, product, method pages)
References
See Help:References for how to manage references in EcoliWiki.