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Sci Rep. 2017 May 3;7(1):1393. doi: 10.1038/s41598-017-00742-3.

Functional organization of protein determinants of meiotic DNA break hotspots.

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Division of Basic Sciences, Fred Hutchinson Cancer Research Center, Seattle, WA, 98109, USA.
Department of Microbiology and Immunology, University of California at San Francisco, San Francisco, CA, 94143, USA.
Instituto de Biología Funcional y Genómica, CSIC/University of Salamanca, C/Zacarías González 2, 37007, Salamanca, Spain.
Division of Basic Sciences, Fred Hutchinson Cancer Research Center, Seattle, WA, 98109, USA.


During Schizosaccharomyces pombe meiotic prophase, homologous chromosomes are co-aligned by linear elements (LinEs) analogous to the axial elements of the synaptonemal complex (SC) in other organisms. LinE proteins also promote the formation of meiotic DNA double-strand breaks (DSBs), the precursors of cross-overs. Rec10 is required for essentially all DSBs and recombination, and three others (Rec25, Rec27, and Mug20) are protein determinants of DSB hotspots - they bind DSB hotspots with high specificity and are required for DSB formation there. These four LinE proteins co-localize in the nucleus in an interdependent way, suggesting they form a complex. We used random mutagenesis to uncover recombination-deficient missense mutants with novel properties. Some missense mutations changed essential residues conserved among Schizosaccharomyces species. DSB formation, gene conversion, and crossing-over were coordinately reduced in the mutants tested. Based on our mutant analysis, we revised the rec27 open reading frame: the new start codon is in the previously annotated first intron. Genetic and fluorescence-microscopy assays indicated that the Rec10 N- and C-terminal regions have complex interactions with Rec25. These mutants are a valuable resource to elucidate further how LinE proteins and the related SCs of other species regulate meiotic DSB formation to form crossovers crucial for meiosis.

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