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Title: Rad54 and Rdh54 occupy spatially and functionally distinct sites within the Rad51-ssDNA presynaptic complex.

Authors: Crickard, J Brooks; Kwon, Youngho; Sung, Patrick; Greene, Eric C

Published In EMBO J, (2020 10 15)

Abstract: Rad54 and Rdh54 are closely related ATP-dependent motor proteins that participate in homologous recombination (HR). During HR, these enzymes functionally interact with the Rad51 presynaptic complex (PSC). Despite their importance, we know little about how they are organized within the PSC, or how their organization affects PSC function. Here, we use single-molecule optical microscopy and genetic analysis of chimeric protein constructs to evaluate the binding distributions of Rad54 and Rdh54 within the PSC. We find that Rad54 and Rdh54 have distinct binding sites within the PSC, which allow these proteins to act cooperatively as DNA sequences are aligned during homology search. Our data also reveal that Rad54 must bind to a specific location within the PSC, whereas Rdh54 retains its function in the repair of MMS-induced DNA damage even when recruited to the incorrect location. These findings support a model in which the relative binding sites of Rad54 and Rdh54 help to define their functions during mitotic HR.

PubMed ID: 32790929 Exiting the NIEHS site

MeSH Terms: Binding Sites; Catalytic Domain/genetics; Cell Cycle Proteins/genetics; Cell Cycle Proteins/metabolism; Chromosome Pairing*; DNA Helicases/genetics; DNA Helicases/metabolism*; DNA Repair Enzymes/genetics; DNA Repair Enzymes/metabolism*; DNA Repair/genetics; DNA Topoisomerases/genetics; DNA Topoisomerases/metabolism*; DNA, Single-Stranded/metabolism*; DNA-Binding Proteins/metabolism; Mutation; Protein Binding; Protein Domains; Rad51 Recombinase/genetics; Rad51 Recombinase/metabolism*; Recombinant Proteins; Saccharomyces cerevisiae Proteins/genetics; Saccharomyces cerevisiae Proteins/metabolism*; Saccharomyces cerevisiae/metabolism*

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