Rrp1 translocase and ubiquitin ligase activities restrict the genome destabilising effects of Rad51 in fission yeast

Nucleic Acids Res. 2021 Jul 9;49(12):6832-6848. doi: 10.1093/nar/gkab511.

Abstract

Rad51 is the key protein in homologous recombination that plays important roles during DNA replication and repair. Auxiliary factors regulate Rad51 activity to facilitate productive recombination, and prevent inappropriate, untimely or excessive events, which could lead to genome instability. Previous genetic analyses identified a function for Rrp1 (a member of the Rad5/16-like group of SWI2/SNF2 translocases) in modulating Rad51 function, shared with the Rad51 mediator Swi5-Sfr1 and the Srs2 anti-recombinase. Here, we show that Rrp1 overproduction alleviates the toxicity associated with excessive Rad51 levels in a manner dependent on Rrp1 ATPase domain. Purified Rrp1 binds to DNA and has a DNA-dependent ATPase activity. Importantly, Rrp1 directly interacts with Rad51 and removes it from double-stranded DNA, confirming that Rrp1 is a translocase capable of modulating Rad51 function. Rrp1 affects Rad51 binding at centromeres. Additionally, we demonstrate in vivo and in vitro that Rrp1 possesses E3 ubiquitin ligase activity with Rad51 as a substrate, suggesting that Rrp1 regulates Rad51 in a multi-tiered fashion.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adenosine Triphosphatases / metabolism
  • DNA, Fungal / metabolism
  • DNA-Binding Proteins / isolation & purification
  • DNA-Binding Proteins / metabolism*
  • DNA-Binding Proteins / physiology
  • Genomic Instability
  • Rad51 Recombinase / metabolism*
  • Schizosaccharomyces / genetics
  • Schizosaccharomyces / metabolism
  • Schizosaccharomyces pombe Proteins / isolation & purification
  • Schizosaccharomyces pombe Proteins / metabolism*
  • Schizosaccharomyces pombe Proteins / physiology
  • Ubiquitin-Protein Ligases / metabolism*

Substances

  • DNA, Fungal
  • DNA-Binding Proteins
  • RHP51 protein, S pombe
  • Rrp1 protein, S pombe
  • Rrp2 protein, S pombe
  • Schizosaccharomyces pombe Proteins
  • Ubiquitin-Protein Ligases
  • Rad51 Recombinase
  • Adenosine Triphosphatases