期刊
MOLECULAR CELL
卷 64, 期 5, 页码 926-939出版社
CELL PRESS
DOI: 10.1016/j.molcel.2016.10.020
关键词
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资金
- Sir Henry Wellcome Postdoctoral Fellowship from the Wellcome Trust [110014/Z/15/Z]
- Francis Crick Institute
- Cancer Research UK [FC0010048]
- UK Medical Research Council [FC0010048]
- Wellcome Trust [FC0010048]
- European Research Council (ERC) Advanced Investigator grant (RecMitMei)
- Wellcome Trust Senior Investigator grant
- Czech Science Foundation [GA13-26629S, GAP207/12/2323]
- National Program of Sustainability II (MEYS CR) [LQ1605]
- project FNUSA-ICRC [CZ.1.05/1.1.00/02.0123]
- project ICRC-ERA-HumanBridge - European Commission [316345]
- NIH [1R35GM118026]
- Cancer Research UK [11581] Funding Source: researchfish
- The Francis Crick Institute [10004] Funding Source: researchfish
- The Francis Crick Institute
- Cancer Research UK [10048] Funding Source: researchfish
- Wellcome Trust [110014/Z/15/Z] Funding Source: Wellcome Trust
Central to homologous recombination in eukaryotes is the RAD51 recombinase, which forms helical nucleoprotein filaments on single-stranded DNA (ssDNA) and catalyzes strand invasion with homologous duplex DNA. Various regulatory proteins assist this reaction including the RAD51 paralogs. We recently discovered that a RAD51 paralog complex from C. elegans, RFS-1/RIP-1, functions predominantly downstream of filament assembly by binding and remodeling RAD-51-ssDNA filaments to a conformation more proficient for strand exchange. Here, we demonstrate that RFS-1/RIP-1 acts by shutting down RAD-51 dissociation from ssDNA. Using stopped-flow experiments, we show that RFS-1/RIP-1 confers this dramatic stabilization by capping the 50 end of RAD-51-ssDNA filaments. Filament end capping propagates a stabilizing effect with a 5'-> 3' polarity approximately 40 nucleotides along individual filaments. Finally, we discover that filament capping and stabilization are dependent on nucleotide binding, but not hydrolysis by RFS-1/RIP-1. These data define the mechanism of RAD51 filament remodeling by RAD51 paralogs.
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