the radially swollen 4 separase mutation of arabidopsis thaliana blocks chromosome disjunction and disrupts the radial microtubule system in meiocytes拟南芥块的径向膨胀4 separase突变染色体分离和扰乱了径向性母细胞微管系统.pdfVIP

The Radially Swollen 4 Separase Mutation of Arabidopsis thaliana Blocks Chromosome Disjunction and Disrupts the Radial Microtubule System in Meiocytes 1 1 1 2 2 1 Xiaohui Yang , Kingsley A. Boateng , Li Yuan , Shuang Wu , Tobias I. Baskin , Christopher A. Makaroff * 1 Department of Chemistry and Biochemistry, Miami University, Oxford, Ohio, United States of America, 2 Biology Department, University of Massachusetts Amherst, Amherst, Massachusetts, United States of America Abstract The caspase-family protease, separase, is required at the onset of anaphase to cleave the cohesin complex that joins replicated sister chromatids. However, in various eukaryotes, separase has acquired additional and distinct functions. A single amino-acid substitution in separase is responsible for phenotypes of the Arabidopsis thaliana mutant, radially swollen 4 (rsw4). This is a conditional mutant, resembling the wild type at the permissive temperature (,20uC) and expressing mutant phenotypes at the restrictive temperature (,30uC). Root cells in rsw4 at the restrictive temperature undergo non- disjunction and other indications of the loss of separase function. To determine to what extent separase activity remains at 30uC, we examined the effect of the mutation on meiosis, where the effects of loss of separase activity through RNA interference are known; and in addition, we examined female gametophyte development. Here, we report that, at the restrictive temperature, replicated chromosomes in rsw4 meiocytes typically fail to disjoin and the cohesin complex remains at centromeres after metaphase. Meiotic spindles appear normal in rsw4 male meiocytes; however the