Mutation of the ATPase domain of MutS homolog-5 (MSH5) reveals a requirement for a functional MutSγ complex for all crossovers in mammalian meiosis

Carolyn R. Milano, J. Kim Holloway, Yongwei Zhang, Bo Jin, Aviv Bergman, Winfried Edelmann, Paula E. Cohen

Research output: Contribution to journalArticlepeer-review

Abstract

During meiosis, induction of DNA double strand breaks (DSB) leads to recombination between homologous chromosomes, resulting in crossovers (CO) and non-crossovers (NCO). Only 10% DSBs resolve as COs, mostly through a class I pathway dependent on MutSγ (MSH4/ MSH5). Class II CO events represent a minor proportion of the total CO count and also arise from DSBs, but are not thought to involve MutSγ. However, loading of MutSγ occurs very early in prophase I at a frequency that far exceeds the final number of class I COs found in late prophase I. Moreover, loss of MutSγ in mouse results in apoptosis before CO formation, preventing analysis of its CO function. We generated a mutation in the ATP binding domain of Msh5 (Msh5GA). While this mutation was not expected to affect MutSγ complex formation, MutSγ foci do not accumulate during prophase I. Nevertheless, while some spermatocytes from Msh5-/- animals progress into pachynema, most spermatocytes from Msh5GA/GA mice progress to late pachynema and beyond. Some spermatocytes from Msh5GA/GA mice complete prophase I entirely, allowing for the first time an assessment of MSH5 function in CO formation. At pachynema, Msh5GA/GA spermatocytes show persistent DSBs, incomplete homolog pairing, and fail to accumulate MutLγ (MLH1/MLH3). Unexpectedly, Msh5GA/GA diakinesis-staged spermatocytes have no chiasmata at all from any CO pathway, indicating that a functional MutSγ complex in early prophase I is a pre-requisite for all COs. ARTICLE SUMMARY MSH4/MSH5 are critical components of the class I crossover (CO) machinery, which is responsible for >90% of the COs that arise in mammalian meiosis. We generated a point mutation in the ATP binding motif of Msh5, and found that mutant spermatocytes lose all COs, not just those arising from the class I pathway.

Original languageEnglish (US)
JournalUnknown Journal
DOIs
StatePublished - Feb 10 2019

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)
  • Agricultural and Biological Sciences(all)
  • Immunology and Microbiology(all)
  • Neuroscience(all)
  • Pharmacology, Toxicology and Pharmaceutics(all)

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