RAD51AP1 mediates RAD51 activity through nucleosome interaction

J Biol Chem. 2021 Jul;297(1):100844. doi: 10.1016/j.jbc.2021.100844. Epub 2021 May 28.


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.

Keywords: RAD51; RAD51AP1; chromatin; homologous recombination DNA repair; mononucleosomes; synaptic complex assembly.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Chromatin / chemistry
  • Chromatin / genetics*
  • Chromosome Pairing / genetics
  • DNA / chemistry
  • DNA / genetics
  • DNA-Binding Proteins / chemistry
  • DNA-Binding Proteins / genetics*
  • Genomic Instability / genetics
  • Histones / chemistry
  • Histones / genetics
  • Humans
  • Nucleosomes / genetics
  • Protein Domains / genetics
  • RNA-Binding Proteins / chemistry
  • RNA-Binding Proteins / genetics*
  • Rad51 Recombinase / chemistry
  • Rad51 Recombinase / genetics*
  • Recombinational DNA Repair / genetics*
  • Telomere / genetics


  • Chromatin
  • DNA-Binding Proteins
  • Histones
  • Nucleosomes
  • RAD51AP1 protein, human
  • RNA-Binding Proteins
  • DNA
  • Rad51 Recombinase