A Single-Strand Annealing Protein Clamps DNA to Detect and Secure Homology

作者：Ander Marcel; Subramaniam Sivaraman; Fahmy Karim; Stewart A Francis^*; Schaeffer Erik

来源：PLOS BIOLOGY, 2015, 13(8): e1002213.

DOI：10.1371/journal.pbio.1002213

Repair of DNA breaks by single-strand annealing (SSA) is a major mechanism for the maintenance of genomic integrity. SSA is promoted by proteins (single-strand-annealing proteins [SSAPs]), such as eukaryotic RAD52 and lambda phage Red beta. These proteins use a short single-stranded region to find sequence identity and initiate homologous recombination. However, it is unclear how SSAPs detect homology and catalyze annealing. Using single-molecule experiments, we provide evidence that homology is recognized by Red beta monomers that weakly hold single DNA strands together. Once annealing begins, dimerization of Red beta clamps the double-stranded region and nucleates nucleoprotein filament growth. In this manner, DNA clamping ensures and secures a successful detection for DNA sequence homology. The clamp is characterized by a structural change of Red beta and a remarkable stability against force up to 200 pN. Our findings not only present a detailed explanation for SSAP action but also identify the DNA clamp as a very stable, noncovalent, DNA-protein interaction.

• 出版日期2015-8