Nature 385, 176 - 181 (09 January 1997); doi:10.1038/385176a0

Structure of the single-stranded-DNA-binding domain of replication protein A bound to DNA

Alexey Bochkarev, Richard A. Pfuetzner, Aled M. Edwards & Lori Frappier

Institute for Molecular Biology and Biotechnology, Cancer Research Group, McMaster University, 1200 Main St. W., Hamilton, Ontario L8N 3Z5, Canada

THE single-stranded-DNA-binding proteins (SSBs) are essential for DNA function in prokaryotic and eukaryotic cells, mitochondria, phages and viruses^1,2. The structures of four SSBs have been solved^3–7, but the molecular details of the interaction of SSBs with DNA remain speculative. We report here the crystal structure at 2.4 Å resolution of the single-stranded-DNA-binding domain of human replication protein A (RPA) bound to DNA. Replication protein A is a heterotrimeric SSB that is highly conserved in eukaryotes. The largest subunit, RPA70, binds to single-stranded (ss)DNA^8,9 and mediates interactions with many cellular and viral proteins¹⁰. The DNA-binding domain, which lies in the middle of RPA70, comprises two structurally homologous sub-domains oriented in tandem. The ssDNA lies in a channel that extends from one subdomain to the other. The structure of each RPA70 subdomain is similar to those of the bacteriophage SSBs, indicating that the mechanism of ssDNA-binding is conserved.

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