Cooper Katrina F, Mallory Michael J, Guacci Vincent, Lowe Katherine, Strich Randy
Department of Biochemistry, Drexel University College of Medicine, Philadelphia, PA 19102, USA.
Genetics. 2009 Jan;181(1):65-79. doi: 10.1534/genetics.108.095513. Epub 2008 Nov 10.
Sister-chromatid separation at the metaphase-anaphase transition is regulated by a proteolytic cascade. Destruction of the securin Pds1p liberates the Esp1p separase, which ultimately targets the mitotic cohesin Mcd1p/Scc1p for destruction. Pds1p stabilization by the spindle or DNA damage checkpoints prevents sister-chromatid separation while mutants lacking PDS1 (pds1Delta) are temperature sensitive for growth due to elevated chromosome loss. This report examined the role of the budding yeast Pds1p in meiotic progression using genetic, cytological, and biochemical assays. Similar to its mitotic function, Pds1p destruction is required for metaphase I-anaphase I transition. However, even at the permissive temperature for growth, pds1Delta mutants arrest with prophase I spindle and nuclear characteristics. This arrest was partially suppressed by preventing recombination initiation or by inactivating a subset of recombination checkpoint components. Further studies revealed that Pds1p is required for recombination in both double-strand-break formation and synaptonemal complex assembly. Although deleting PDS1 did not affect the degradation of the meiotic cohesin Rec8p, Mcd1p was precociously destroyed as cells entered the meiotic program. This role is meiosis specific as Mcd1p destruction is not altered in vegetative pds1Delta cultures. These results define a previously undescribed role for Pds1p in cohesin maintenance, recombination, and meiotic progression.
中期-后期转换时姐妹染色单体的分离由一个蛋白水解级联反应调控。securin蛋白Pds1p的降解释放出Esp1p分离酶,后者最终靶向有丝分裂黏连蛋白Mcd1p/Scc1p使其降解。纺锤体或DNA损伤检查点对Pds1p的稳定作用可防止姐妹染色单体分离,而缺乏PDS1的突变体(pds1Δ)由于染色体丢失增加,在生长方面对温度敏感。本报告使用遗传学、细胞学和生化分析方法研究了芽殖酵母Pds1p在减数分裂进程中的作用。与其在有丝分裂中的功能相似,中期I-后期I转换需要Pds1p的降解。然而,即使在允许生长的温度下,pds1Δ突变体也会因前期I纺锤体和细胞核特征而停滞。通过阻止重组起始或使一部分重组检查点成分失活,这种停滞得到了部分抑制。进一步研究表明,Pds1p在双链断裂形成和联会复合体组装的重组过程中都是必需的。虽然删除PDS1并不影响减数分裂黏连蛋白Rec8p的降解,但随着细胞进入减数分裂程序,Mcd1p会过早被破坏。这种作用是减数分裂特有的,因为在营养型pds1Δ培养物中Mcd1p的破坏并未改变。这些结果确定了Pds1p在黏连蛋白维持、重组和减数分裂进程中一个以前未被描述的作用。