RAD51AP1 mediates RAD51 activity through nucleosome interaction
| Autor: | Weixing Zhao, Platon Selemenakis, Bo Wu, Yuxin Huang, Dauren S. Alimbetov, Claudia Wiese, Elena Pires, Neelam Sharma |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2021 |
| Předmět: |
0301 basic medicine
Genome instability EMSA electrophoretic mobility shift assay RAD51 Biochemistry Histones chemistry.chemical_compound PVDF polyvinylidene difluoride mononucleosomes Histone octamer homologous recombination DNA repair synaptic complex assembly ANOVA analysis of variance Chemistry NCP nucleosome core particle RAD51AP1 RNA-Binding Proteins Hop2 homologous pairing 2 Telomere Chromatin Cell biology Nucleosomes DNA-Binding Proteins PALB2 partner and localizer of BRCA2 Research Article MNase micrococcal nuclease NAP1 nucleosome-associated protein 1 ATP adenosine triphosphate SEC size-exclusion chromatography MBP maltose-binding protein Genomic Instability HPLC high-pressure liquid chromatography 03 medical and health sciences Protein Domains Nucleosome Humans AA amino acid Molecular Biology Mnd1 meiotic nuclear divisions 1 UAF1 USP1-associated factor 1 MMC mitomycin C 030102 biochemistry & molecular biology Recombinational DNA Repair D-loop displacement-loop Cell Biology DNA Repair Pathway DNA-binding domain DNA Ni-NTA nickel nitrilotriacetic acid Nucleoprotein Chromosome Pairing 030104 developmental biology RAD51AP1 RAD51-associated protein 1 BSA bovine serum albumin Rad51 Recombinase HR homologous recombination Homologous recombination |
| Zdroj: | The Journal of Biological Chemistry |
| ISSN: | 1083-351X 0021-9258 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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