Meiotic association between Spo11 regulated by Rec102, Rec104 and Rec114.
Meiotic recombination is initiated by DNA double-stranded break (DSB) formation catalyzed by Spo11, a type-II topoisomerase-like transesterificase, presumably via a dimerization-mediated mechanism. We demonstrate the existence of in vivo interactions between Spo11 proteins carrying distinct tags, and the chromatin-binding and DSB activity of tagged Spo11 at innate and targeted DSB sites upon fusion to the Gal4 DNA-binding domain. First we identified the interaction between Spo11-3FLAG and Gal4BD-Spo11 proteins, and established that this interaction specifically occurs at the time of DSB formation. We then observed that presence of the Gal4BD-spo11Y135F (nuclease-deficient) protein allows Spo11-3FLAG recruitment at the GAL2 locus, indicative of the formation of a hetero-complex near the GAL2 UAS sites, but no formation of double- or single-strand breaks. Spo11 self-interaction around the GAL2 DSB site depends on other proteins for DSB formation, in particular Rec102, Rec104 and Rec114. Together, these results suggest that in vivo self-association of Spo11 during meiosis is genetically regulated. The results are discussed in relation to possible roles of Spo11 self-interaction in the control of the cleavage activity.
Sasanuma
Hiroyuki
H
Genetic System Regulation Laboratory, RIKEN Discovery Research Institute, Hirosawa 2-1, Wako, Saitama 351-0198, Japan.
Murakami
Hajime
H
Fukuda
Tomoyuki
T
Shibata
Takehiko
T
Nicolas
Alain
A
Ohta
Kunihiro
K
D000217:Acyltransferases / Q000235:genetics; D002843:Chromatin / Q000378:metabolism; D053903:DNA Breaks, Double-Stranded; D004250:DNA Topoisomerases, Type II / Q000235:genetics / Q000378:metabolism*; D004268:DNA-Binding Proteins / Q000378:metabolism*; D004706:Endodeoxyribonucleases; D008540:Meiosis; D009004:Monosaccharide Transport Proteins / Q000235:genetics; D009154:Mutation; D009842:Oligopeptides; D010455:Peptides / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D045522:Recombinases; D029701:Saccharomyces cerevisiae Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D014157:Transcription Factors / Q000737:chemistry; D000099257:Meiotic Recombination Protein SPO11
D000217:Acyltransferases; D002843:Chromatin; D004250:DNA Topoisomerases, Type II; D004268:DNA-Binding Proteins; D004706:Endodeoxyribonucleases; D009004:Monosaccharide Transport Proteins; D009842:Oligopeptides; D010455:Peptides; D011993:Recombinant Fusion Proteins; D045522:Recombinases; D029701:Saccharomyces cerevisiae Proteins; D014157:Transcription Factors; C106901:GAL2 protein, S cerevisiae; C052130:GAL4 protein, S cerevisiae; C073372:REC102 protein, S cerevisiae; C080594:REC104 protein, S cerevisiae; C080622:REC114 protein, S cerevisiae; C085706:FLAG peptide; C518938:Spo11 protein, S cerevisiae; D000099257:Meiotic Recombination Protein SPO11