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Interactions of human rad54 protein with branched DNA molecules
Journal article   Open access   Peer reviewed

Interactions of human rad54 protein with branched DNA molecules

Olga M Mazina, Matthew J Rossi, Nicolas H Thomaä and Alexander V Mazin
The Journal of biological chemistry, v 282(29), pp 21068-21080
20 Jul 2007
PMID: 17545145
url
https://doi.org/10.1074/jbc.M701992200View
Published, Version of Record (VoR) Open

Abstract

Protein Structure, Tertiary Cross-Linking Reagents - pharmacology Humans Glutathione Transferase - metabolism Ions Nuclear Proteins - chemistry Hydrolysis DNA - chemistry DNA Helicases Magnesium - chemistry Protein Binding Adenosine Triphosphatases - chemistry DNA, Cruciform Kinetics Adenosine Triphosphate - chemistry Nucleic Acid Conformation DNA-Binding Proteins
The Rad54 protein plays an important role during homologous recombination in eukaryotes. The protein belongs to the Swi2/Snf2 family of ATP-dependent DNA translocases. We previously showed that yeast and human Rad54 (hRad54) specifically bind to Holliday junctions and promote branch migration. Here we examined the minimal DNA structural requirements for optimal hRad54 ATPase and branch migration activity. Although a 12-bp double-stranded DNA region of branched DNA is sufficient to induce ATPase activity, the minimal substrate that gave rise to optimal stimulation of the ATP hydrolysis rate consisted of two short double-stranded DNA arms, 15 bp each, combined with a 45-nucleotide single-stranded DNA branch. We showed that hRad54 binds preferentially to the open and not to the stacked conformation of branched DNA. Stoichiometric titration of hRad54 revealed formation of two types of hRad54 complexes with branched DNA substrates. The first of them, a dimer, is responsible for the ATPase activity of the protein. However, branch migration activity requires a significantly higher stoichiometry of hRad54, approximately 10 +/- 2 protein monomers/DNA molecule. This pleomorphism of hRad54 in formation of oligomeric complexes with DNA may correspond to multiple functions of the protein in homologous recombination.

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Collaboration types
Domestic collaboration
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Web of Science research areas
Biochemistry & Molecular Biology
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