Crystal structure of a Rad51 filament

Abstract

Rad51, the major eukaryotic homologous recombinase, is important for the repair of DNA damage and the maintenance of genomic diversity and stability. The active form of this DNA-dependent ATPase is a helical filament within which the search for homology and strand exchange occurs. Here we present the crystal structure of a Saccharomyces cerevisiae Rad51 filament formed by a gain-of-function mutant. This filament has a longer pitch than that seen in crystals of Rad51's prokaryotic homolog RecA, and places the ATPase site directly at a new interface between protomers. Although the filament exhibits approximate six-fold symmetry, alternate protein-protein interfaces are slightly different, implying that the functional unit of Rad51 within the filament may be a dimer. Additionally, we show that mutation of His352, which lies at this new interface, markedly disrupts DNA binding.

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Figure 1: N-terminally truncated Rad51 catalyzes strand exchange.
Figure 2: Filaments of Rad51 and RecA.
Figure 3: Oligomerization of Rad51.
Figure 4: The Rad51 filament has two types of protomer-protomer interactions at the ATPase site.
Figure 5: Binding to ssDNA (φX174).
Figure 6: Tyrosine phosphorylation site at the protomer-protomer interface.

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Acknowledgements

We thank the staff of BioCARS and of Structural Biology Center for help with data collection. Use of the Advanced Photon Source was supported by the US Department of Energy, Basic Energy Sciences, Office of Science, under contract no. W-31-109-Eng-38. Use of the BioCARS Sector 14 was supported by the US National Institutes of Health (NIH), National Center for Research Resources, under grant RR07707. This work was supported in part by NIH GM54099 (L.S.S.), NIH GM 058827 (P.A.R.), NIH 2 T32 GM008720 (A.B.C.), NIH T32 CA09594 (T.W.L.) and NIH T32 CA09503 (C.F.).

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Correspondence to Phoebe A Rice.

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Supplementary information

Supplementary Figure 1

Representative electron density. (PDF 340 kb)

Supplementary Figure 2

Anomalous electron density. (PDF 98 kb)

Supplementary Figure 3

Early electron density. (PDF 222 kb)

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Conway, A., Lynch, T., Zhang, Y. et al. Crystal structure of a Rad51 filament. Nat Struct Mol Biol 11, 791–796 (2004). https://doi.org/10.1038/nsmb795

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