RPA resolves conflicting activities of accessory proteins during reconstitution of Dmc1-mediated meiotic recombination
| Autor: | Douglas K. Bishop, Benjamin P. Weissman, Yuen-Ling Chan, Annie Zhang |
|---|---|
| Rok vydání: | 2018 |
| Předmět: |
Saccharomyces cerevisiae Proteins
Chromosomal Proteins Non-Histone RAD51 DNA Single-Stranded Cell Cycle Proteins Genome Integrity Repair and Replication 03 medical and health sciences chemistry.chemical_compound 0302 clinical medicine Replication Protein A Genetics Protein secondary structure 030304 developmental biology Recombination Genetic 0303 health sciences biology fungi biology.organism_classification Cell biology Nucleoprotein DNA-Binding Proteins Meiosis enzymes and coenzymes (carbohydrates) Saccharomycetales chemistry Nucleic Acid Conformation DMC1 Rad51 Recombinase Homologous recombination 030217 neurology & neurosurgery Recombination DNA |
| Zdroj: | Nucleic Acids Research |
| ISSN: | 1362-4962 0305-1048 |
| DOI: | 10.1093/nar/gky1160 |
| Popis: | Dmc1 catalyzes homology search and strand exchange during meiotic recombination in budding yeast and many other organisms including humans. Here we reconstitute Dmc1 recombination in vitro using six purified proteins from budding yeast including Dmc1 and its accessory proteins RPA, Rad51, Rdh54/Tid1, Mei5-Sae3 and Hop2-Mnd1 to promote D-loop formation between ssDNA and dsDNA substrates. Each accessory protein contributed to Dmc1’s activity, with the combination of all six proteins yielding optimal activity. The ssDNA binding protein RPA plays multiple roles in stimulating Dmc1’s activity including by overcoming inhibitory effects of ssDNA secondary structure on D-loop reactions, and by elongating D-loops. In addition, we demonstrate that RPA limits inhibitory interactions of Hop2-Mnd1 and Rdh54/Tid1 that otherwise occur during assembly of Dmc1-ssDNA nucleoprotein filaments. Finally, we report interactions between the proteins employed in the biochemical reconstitution including a direct interaction between Rad51 and Dmc1 that is enhanced by Mei5-Sae3. |
| Databáze: | OpenAIRE |
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