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Catalysis of homologous DNA pairing by yeast Rad51 and Rad54 proteins

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Abstract

The Saccharomyces cerevisiae RAD51 and RAD54 genes are both required for the occurrence of homologous recombination and for the repair of double-stranded DNA breaks1. Previous studies have indicated that Rad51 protein, together with the single-stranded DNA-binding factor replication protein A (RPA), can promote the formation of heteroduplex DNA2,3,4, which is a key intermediate in homologous recombination1. Here we report the purification of the Rad54 protein to near homogeneity and the biochemical testing of its molecular function. We find that Rad54 protein possesses a double-stranded DNA-dependent ATPase activity, and that it interacts with the Rad51 protein. Addition of Rad54 protein to reactions containing Rad51 strongly stimulates the rate of pairing between homologous single-stranded and double-stranded DNA molecules. We conclude that Rad54 acts to overcome kinetic impediments that would limit homologous DNA pairing between recombining chromosomes in vivo.

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Figure 1: Overexpression and purification of Rad54 protein.
Figure 2: Rad54 protein has DNA-dependent ATPase activity.
Figure 3: Rad54 functions with Rad51 to promote homologous DNA pairing.
Figure 4: D-loop formation requires Rad54 protein.

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Acknowledgements

This work was supported by a PHS grant from the National Institute of Environmental Health Sciences.

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Authors and Affiliations

  1. Institute of Biotechnology and Department of Molecular Medicine, University of Texas Health Science Center at San Antonio, 15355 Lambda Drive, San Antonio, 78245, Texas, USA

    Galina Petukhova, Sabrina Stratton & Patrick Sung

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  1. Galina Petukhova
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  2. Sabrina Stratton
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  3. Patrick Sung
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Correspondence to Patrick Sung.

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Petukhova, G., Stratton, S. & Sung, P. Catalysis of homologous DNA pairing by yeast Rad51 and Rad54 proteins. Nature 393, 91–94 (1998). https://doi.org/10.1038/30037

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