Proteomic analysis of the Ade6-M26 meiotic recombination hotspot in the fission yeast schizosaccharomyces pombe

ABSTRACT: Homologous recombination events during meiosis cluster at specific loci called hotspots. Hotspot activation requires accessible chromatin, DNA sequence motifs DNA sequence-specific binding proteins, but how these proteins create a suitable environment for Rec12 recruitment is unknown. Here, we purified the ade6-M26 meiotic recombination hotspot from Schizosaccharomyces pombe and identified novel regulators of hotspot activation by mass spectrometry. Small, circular minichromosomes containing the ade6-M26 hotspot or control allele were transformed into strains permitting synchronous meiotic induction and purified from these cultures at sequential time points. Enrichment of known M26 binding factors Atf1 and Pcr1 in the data set validated the approach. Filtering for chromatin-related proteins enriched in the hotspot revealed candidate hotspot regulators, which were validated using genetic studies. This study highlights the power of unbiased proteomic screens to reveal novel insights into fundamental biological processes, and highlights a critical role in H2AZ turnover in mediating access of transcription factors to their underlying DNA-sequence motifs.

INSTRUMENT(S): Orbitrap Fusion

ORGANISM(S): Schizosaccharomyces Pombe

SUBMITTER: Aaron Storey  

LAB HEAD: Wayne P Wahls

PROVIDER: PXD009649 | Pride | 2018-11-04

REPOSITORIES: Pride