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Rad54 protein promotes branch migration of Holliday junctions

Nature volume 442, pages 590–593 (2006)Cite this article

Abstract

Homologous recombination has a crucial function in the repair of DNA double-strand breaks and in faithful chromosome segregation1,2,3. The mechanism of homologous recombination involves the search for homology and invasion of the ends of a broken DNA molecule into homologous duplex DNA to form a cross-stranded structure, a Holliday junction (HJ)4,5,6,7. A HJ is able to undergo branch migration along DNA, generating increasing or decreasing lengths of heteroduplex. In both prokaryotes and eukaryotes, the physical evidence for HJs, the key intermediate in homologous recombination, was provided by electron microscopy8. In bacteria there are specialized enzymes that promote branch migration of HJs7. However, in eukaryotes the identity of homologous recombination branch-migration protein(s) has remained elusive. Here we show that Rad54, a Swi2/Snf2 protein9, binds HJ-like structures with high specificity and promotes their bidirectional branch migration in an ATPase-dependent manner. The activity seemed to be conserved in human and yeast Rad54 orthologues. In vitro, Rad54 has been shown to stimulate DNA pairing of Rad51, a key homologous recombination protein10,11,12. However, genetic data indicate that Rad54 protein might also act at later stages of homologous recombination, after Rad51 (ref. 13). Novel DNA branch-migration activity is fully consistent with this late homologous recombination function of Rad54 protein.

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Figure 1: hRad54 protein binds preferentially to branched DNA molecules.
Figure 2: Rad54 promotes an ATPase-dependent BM of synthetic PX junction DNA.
Figure 3: Rad54 promotes BM of synthetic four-way HJs (X junctions).
Figure 4: hRad54 protein catalyses BM of joint molecules (α-structures) generated by hRad51 in four-stranded DNA exchange.

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Acknowledgements

We thank P. Sung and M. Wold for hRad51 and hRPA expression vectors; S. Kowalczykowski, M. Spies and A. Alexeev for RecA, yRad51 and yRad54; J. Kadonaga for ISWI; G. Schnitzler and N. Ulyanova for hSWI/SNF; M. Bouchard and A. Kuzminov for comments and discussion. This work was supported by an NIH grant to A.V.M.

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Authors and Affiliations

  1. Department of Biochemistry and Molecular Biology, Drexel University College of Medicine, Pennsylvania, 19102-1192, Philadelphia, USA

    Dmitry V. Bugreev, Olga M. Mazina & Alexander V. Mazin

  2. Institute of Chemical Biology and Fundamental Medicine, Siberian Branch of Russian Academy of Science, 630090, Novosibirsk, Russia

    Dmitry V. Bugreev

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Correspondence to Alexander V. Mazin.

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Bugreev, D., Mazina, O. & Mazin, A. Rad54 protein promotes branch migration of Holliday junctions. Nature 442, 590–593 (2006). https://doi.org/10.1038/nature04889

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