RAD51AP1 mediates RAD51 activity through nucleosome interaction |
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Authors: | Elena Pires Neelam Sharma Platon Selemenakis Bo Wu Yuxin Huang Dauren S. Alimbetov Weixing Zhao Claudia Wiese |
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Affiliation: | 1.Department of Environmental and Radiological Health Sciences, Colorado State University, Fort Collins, Colorado, USA;2.Cell and Molecular Biology Graduate Program, Colorado State University, Fort Collins, Colorado, USA;3.Department of Biochemistry and Structural Biology, University of Texas Health San Antonio, San Antonio, Texas, USA |
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Abstract: | RAD51-associated protein 1 (RAD51AP1) is a key protein in the homologous recombination (HR) DNA repair pathway. Loss of RAD51AP1 leads to defective HR, genome instability, and telomere erosion. RAD51AP1 physically interacts with the RAD51 recombinase and promotes RAD51-mediated capture of donor DNA, synaptic complex assembly, and displacement-loop formation when tested with nucleosome-free DNA substrates. In cells, however, DNA is packaged into chromatin, posing an additional barrier to the complexities of the HR reaction. In this study, we show that RAD51AP1 binds to nucleosome core particles (NCPs), the minimum basic unit of chromatin in which approximately two superhelical turns of 147 bp double-stranded DNA are wrapped around one histone octamer with no free DNA ends remaining. We identified a C-terminal region in RAD51AP1, including its previously mapped DNA-binding domain, as critical for mediating the association between RAD51AP1 and both the NCP and the histone octamer. Using in vitro surrogate assays of HR activity, we show that RAD51AP1 is capable of promoting duplex DNA capture and initiating joint-molecule formation with the NCP and chromatinized template DNA, respectively. Together, our results suggest that RAD51AP1 directly assists in the RAD51-mediated search for donor DNA in chromatin. We present a model, in which RAD51AP1 anchors the DNA template through affinity for its nucleosomes to the RAD51-ssDNA nucleoprotein filament. |
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Keywords: | homologous recombination DNA repair chromatin mononucleosomes synaptic complex assembly RAD51AP1 RAD51 |
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