PMID:27867009

Citation	Taylor, MRG, Špírek, M, Jian Ma, C, Carzaniga, R, Takaki, T, Collinson, LM, Greene, EC, Krejci, L and Boulton, SJ (2016) A Polar and Nucleotide-Dependent Mechanism of Action for RAD51 Paralogs in RAD51 Filament Remodeling. Mol. Cell 64:926-939
Abstract	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 5' 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.
Links	PubMed PMC5145814 Online version:10.1016/j.molcel.2016.10.020
Keywords	Caenorhabditis elegans Proteins/genetics; Caenorhabditis elegans Proteins/metabolism; Carrier Proteins/genetics; Carrier Proteins/metabolism; DNA, Single-Stranded/genetics; DNA-Binding Proteins/genetics; DNA-Binding Proteins/metabolism; Intermediate Filaments/genetics; Intermediate Filaments/metabolism; Multiprotein Complexes/metabolism; Protein Binding; Rad51 Recombinase/genetics; Rad51 Recombinase/metabolism; Recombinational DNA Repair
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Annotations

Gene product	Qualifier	GO Term	Evidence Code	with/from	Aspect	Extension	Notes	Status
CAEEL:RFS1		GO:0000730: DNA recombinase assembly	ECO:0000316:	UniProtKB:Q21621	P		Figure 1 depicts the cessation of the dissociation of RAD-51 from ssDNA due to the presence of protein RFS-1/RIP-1. Figure 1, Part H [(i), (ii)] shows the known mechanism of RAD-51 by demonstrating that it’s presence prohibits the binding of ScRPA-eGFP-ssDNA. The incapability of RAD-51 to be expressed due to the lack of ATP is found in Figure 1, Part H (ii). Figure 1 ultimately illustrates that in the precence of the protein RFS-1/RIP-1 and ATP; RAD-51 along with the protein RFS-1/RIP-1 is present but the ScRPA-eGFP-ssDNA are not [(Figure 1, Part H (iii)]. The same events occur when the protein RFS-1/RIP-1 is present but not ATP [(Figure 1, Part H (iv)].	complete CACAO 13210

References

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