Role of the Rad52 amino-terminal DNA binding activity in DNA strand capture in homologous recombination
| Autor: | Patrick Sung, Rodney Rothstein, Changhyun Seong, Swee Chuang Lim Hallwyl, Uffe Hasbro Mortensen, Idina Shi |
|---|---|
| Rok vydání: | 2009 |
| Předmět: |
Chromatin Immunoprecipitation
Saccharomyces cerevisiae Proteins HMG-box DNA Repair DNA polymerase II genetic processes DNA Single-Stranded Saccharomyces cerevisiae Biochemistry DNA Breaks Double-Stranded Molecular Biology Replication protein A chemistry.chemical_classification Recombination Genetic DNA ligase DNA clamp Binding Sites biology fungi Cell Biology Molecular biology Branch migration Cell biology Rad52 DNA Repair and Recombination Protein enzymes and coenzymes (carbohydrates) Microscopy Electron chemistry Amino Acid Substitution Mutation DNA: Replication Repair Recombination and Chromosome Dynamics biology.protein DNA supercoil Electrophoresis Polyacrylamide Gel Rad51 Recombinase In vitro recombination Protein Binding |
| Zdroj: | The Journal of biological chemistry. 284(48) |
| ISSN: | 1083-351X |
| Popis: | Saccharomyces cerevisiae Rad52 protein promotes homologous recombination by nucleating the Rad51 recombinase onto replication protein A-coated single-stranded DNA strands and also by directly annealing such strands. We show that the purified rad52-R70A mutant protein, with a compromised amino-terminal DNA binding domain, is capable of Rad51 delivery to DNA but is deficient in DNA annealing. Results from chromatin immunoprecipitation experiments find that rad52-R70A associates with DNA double-strand breaks and promotes recruitment of Rad51 as efficiently as wild-type Rad52. Analysis of gene conversion intermediates reveals that rad52-R70A cells can mediate DNA strand invasion but are unable to complete the recombination event. These results provide evidence that DNA binding by the evolutionarily conserved amino terminus of Rad52 is needed for the capture of the second DNA end during homologous recombination. |
| Databáze: | OpenAIRE |
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