S. cerevisiae Srs2 helicase ensures normal recombination intermediate metabolism during meiosis and prevents accumulation of Rad51 aggregates

Chromosoma. 2019 Sep;128(3):249-265. doi: 10.1007/s00412-019-00705-9. Epub 2019 May 9.

Abstract

We investigated the meiotic role of Srs2, a multi-functional DNA helicase/translocase that destabilises Rad51-DNA filaments and is thought to regulate strand invasion and prevent hyper-recombination during the mitotic cell cycle. We find that Srs2 activity is required for normal meiotic progression and spore viability. A significant fraction of srs2 mutant cells progress through both meiotic divisions without separating the bulk of their chromatin, although in such cells sister centromeres often separate. Undivided nuclei contain aggregates of Rad51 colocalised with the ssDNA-binding protein RPA, suggesting the presence of persistent single-strand DNA. Rad51 aggregate formation requires Spo11-induced DSBs, Rad51 strand-invasion activity and progression past the pachytene stage of meiosis, but not the DSB end-resection or the bias towards interhomologue strand invasion characteristic of normal meiosis. srs2 mutants also display altered meiotic recombination intermediate metabolism, revealed by defects in the formation of stable joint molecules. We suggest that Srs2, by limiting Rad51 accumulation on DNA, prevents the formation of aberrant recombination intermediates that otherwise would persist and interfere with normal chromosome segregation and nuclear division.

Keywords: Budding yeast; Meiosis; Rad51; Recombination; Srs2 helicase.

Publication types

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

MeSH terms

  • DNA Helicases / genetics*
  • DNA Helicases / metabolism
  • MAP Kinase Kinase 1 / metabolism
  • Meiosis*
  • Microbial Viability / genetics
  • Mutation
  • Protein Aggregates
  • Protein Binding
  • Rad51 Recombinase / metabolism*
  • Recombination, Genetic*
  • Saccharomyces cerevisiae / genetics*
  • Saccharomyces cerevisiae / metabolism*
  • Saccharomyces cerevisiae Proteins / genetics*
  • Saccharomyces cerevisiae Proteins / metabolism
  • Spores, Fungal

Substances

  • Protein Aggregates
  • Saccharomyces cerevisiae Proteins
  • SRS2 protein, S cerevisiae
  • MAP Kinase Kinase 1
  • Rad51 Recombinase
  • DNA Helicases