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A novel function of Rad54 protein. Stabilization of the Rad51 nucleoprotein filament
Journal article   Open access   Peer reviewed

A novel function of Rad54 protein. Stabilization of the Rad51 nucleoprotein filament

Alexander V Mazin, Andrei A Alexeev and Stephen C Kowalczykowski
The Journal of biological chemistry, v 278(16), pp 14029-14036
18 Apr 2003
DOI: https://doi.org/10.1074/jbc.M212779200
PMID: 12566442
Featured in Collection :   UN Sustainable Development Goals @ Drexel
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https://doi.org/10.1074/jbc.M212779200View
Published, Version of Record (VoR) Open

Abstract

DNA Restriction Enzymes - metabolism Sodium Chloride - pharmacology Polystyrenes - chemistry DNA-Binding Proteins - metabolism Dose-Response Relationship, Drug Saccharomyces cerevisiae - metabolism Salts - pharmacology Recombination, Genetic Protein Structure, Tertiary Biotinylation Rad51 Recombinase Electrophoresis, Polyacrylamide Gel Adenosine Triphosphatases - metabolism DNA - metabolism DNA Repair Enzymes DNA-Binding Proteins - chemistry DNA, Single-Stranded - chemistry Protein Transport DNA Helicases Streptavidin - metabolism Models, Biological Nucleoproteins - metabolism Saccharomyces cerevisiae Proteins - metabolism Protein Binding Saccharomyces cerevisiae Proteins - physiology
Homologous recombination is important for the repair of double-stranded DNA breaks in all organisms. Rad51 and Rad54 proteins are two key components of the homologous recombination machinery in eukaryotes. In vitro, Rad51 protein assembles with single-stranded DNA to form the helical nucleoprotein filament that promotes DNA strand exchange, a basic step of homologous recombination. Rad54 protein interacts with this Rad51 nucleoprotein filament and stimulates its DNA pairing activity, suggesting that Rad54 protein is a component of the nucleoprotein complex involved in the DNA homology search. Here, using physical criteria, we demonstrate directly the formation of Rad54-Rad51-DNA nucleoprotein co-complexes that contain equimolar amounts of each protein. The binding of Rad54 protein significantly stabilizes the Rad51 nucleoprotein filament formed on either single-stranded DNA or double-stranded DNA. The Rad54-stabilized nucleoprotein filament is more competent in DNA strand exchange and acts over a broader range of solution conditions. Thus, the co-assembly of an interacting partner with the Rad51 nucleoprotein filament represents a novel means of stabilizing the biochemical entity central to homologous recombination, and reveals a new function of Rad54 protein.

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Web of Science research areas
Biochemistry & Molecular Biology
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