title: Identification of Putative Mek1 Substrates during Meiosis in Saccharomyces cerevisiae Using Quantitative Phosphoproteomics.
abstract: Meiotic recombination plays a key role in sexual reproduction as it generates crossovers that, in combination with sister chromatid cohesion, physically connect homologous chromosomes, thereby promoting their proper segregation at the first meiotic division. Meiotic recombination is initiated by programmed double strand breaks (DSBs) catalyzed by the evolutionarily conserved, topoisomerase-like protein Spo11. Repair of these DSBs is highly regulated to create crossovers between homologs that are distributed throughout the genome. This repair requires the presence of the mitotic recombinase, Rad51, as well as the strand exchange activity of the meiosis-specific recombinase, Dmc1. A key regulator of meiotic DSB repair in Saccharomyces cerevisiae is the meiosis-specific kinase Mek1, which promotes interhomolog strand invasion and is required for the meiotic recombination checkpoint and the crossover/noncrossover decision. Understanding how Mek1 regulates meiotic recombination requires the identification of its substrates. Towards that end, an unbiased phosphoproteomic approach utilizing Stable Isotope Labeling by Amino Acids in Cells (SILAC) was utilized to generate a list of potential Mek1 substrates, as well as proteins containing consensus phosphorylation sites for cyclin-dependent kinase, the checkpoint kinases, Mec1/Tel1, and the polo-like kinase, Cdc5. These experiments represent the first global phosphoproteomic dataset for proteins in meiotic budding yeast.
authors.affiliation: Biochemistry and Cell Biology, Stony Brook University, Stony Brook, New York, 11794-5215, United States of America.
authors.forename: Raymond T
authors.initials: RT
authors.lastname: Suhandynata
authors.affiliation: Biochemistry and Cell Biology, Stony Brook University, Stony Brook, New York, 11794-5215, United States of America.
authors.forename: Lihong
authors.initials: L
authors.lastname: Wan
authors.affiliation: Ludwig Institute for Cancer Research, University of California San Diego, La Jolla, California, 92093, United States of America.|Cellular and Molecular Medicine, University of California San Diego, La Jolla, California, 92093, United States of America.
authors.forename: Huilin
authors.initials: H
authors.lastname: Zhou
authors.affiliation: Biochemistry and Cell Biology, Stony Brook University, Stony Brook, New York, 11794-5215, United States of America.
authors.forename: Nancy M
authors.initials: NM
authors.lastname: Hollingsworth
mesh_terms: D001665:Binding Sites; D003434:Crossing Over, Genetic; D053903:DNA Breaks, Double-Stranded; D059765:Homologous Recombination; D007553:Isotope Labeling / Q000379:methods; D048369:MAP Kinase Kinase 1 / Q000737:chemistry / Q000378:metabolism*; D008540:Meiosis; D010750:Phosphoproteins / Q000032:analysis* / Q000378:metabolism; D040901:Proteomics / Q000379:methods*; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000502:physiology*; D029701:Saccharomyces cerevisiae Proteins / Q000737:chemistry / Q000378:metabolism
chemical_list: D010750:Phosphoproteins; D029701:Saccharomyces cerevisiae Proteins; D048369:MAP Kinase Kinase 1