Functional interactions between SPO11 and REC102 during initiation of meiotic recombination in Saccharomyces cerevisiae.
In Saccharomyces cerevisiae, formation of the DNA double-strand breaks (DSBs) that initiate meiotic recombination requires the products of at least 10 genes. Spo11p is thought to be the catalytic subunit of the DNA cleaving activity, but the roles of the other proteins, and the interactions among them, are not well understood. This study demonstrates genetic and physical interactions between the products of SPO11 and another early meiotic gene required for DSB formation, REC102. We found that epitope-tagged versions of SPO11 and REC102 that by themselves were capable of supporting normal or nearly normal levels of meiotic recombination conferred a severe synthetic cold-sensitive phenotype when combined in the same cells. DSB formation, meiotic gene conversion, and spore viability were drastically reduced in the doubly tagged strain at a nonpermissive temperature. This conditional defect could be partially rescued by expression of untagged SPO11, but not by expression of untagged REC102, indicating that tagged REC102 is fully dominant for this synthetic phenotype. Both tagged and wild-type Spo11p co-immunoprecipitated with tagged Rec102p from meiotic cell extracts, indicating that these proteins are present in a common complex in vivo. Tagged Rec102p localized to the nucleus in whole cells and to chromatin on spread meiotic chromosomes. Our results are consistent with the idea that a multiprotein complex that includes Spo11p and Rec102p promotes meiotic DSB formation.
Kee
Kehkooi
K
Molecular Biology Program, Memorial Sloan-Kettering Cancer Center and Weill Graduate School of Medical Sciences of Cornell University, New York, New York 10021, USA.
Keeney
Scott
S
D000345:Affinity Labels; D002843:Chromatin / Q000502:physiology; D003080:Cold Temperature; D004268:DNA-Binding Proteins; D004706:Endodeoxyribonucleases; D004950:Esterases / Q000235:genetics / Q000502:physiology*; D005656:Fungal Proteins / Q000235:genetics / Q000502:physiology*; D008540:Meiosis / Q000502:physiology*; D009687:Nuclear Proteins / Q000502:physiology; D010641:Phenotype; D011233:Precipitin Tests; D045522:Recombinases; D011995:Recombination, Genetic; D012441:Saccharomyces cerevisiae / Q000166:cytology / Q000235:genetics*; D029701:Saccharomyces cerevisiae Proteins; D000099257:Meiotic Recombination Protein SPO11
D000345:Affinity Labels; D002843:Chromatin; D004268:DNA-Binding Proteins; D004706:Endodeoxyribonucleases; D004950:Esterases; D005656:Fungal Proteins; D009687:Nuclear Proteins; D045522:Recombinases; D029701:Saccharomyces cerevisiae Proteins; D000099257:Meiotic Recombination Protein SPO11; C073372:REC102 protein, S cerevisiae