Urban Evelin, Nagarkar-Jaiswal Sonal, Lehner Christian F, Heidmann Stefan K
Lehrstuhl für Genetik, University of Bayreuth, Bayreuth, Germany.
Institute of Molecular Life Sciences (IMLS), University of Zurich, Zurich, Switzerland.
PLoS Genet. 2014 Aug 7;10(8):e1004540. doi: 10.1371/journal.pgen.1004540. eCollection 2014 Aug.
Replicated sister chromatids are held in close association from the time of their synthesis until their separation during the next mitosis. This association is mediated by the ring-shaped cohesin complex that appears to embrace the sister chromatids. Upon proteolytic cleavage of the α-kleisin cohesin subunit at the metaphase-to-anaphase transition by separase, sister chromatids are separated and segregated onto the daughter nuclei. The more complex segregation of chromosomes during meiosis is thought to depend on the replacement of the mitotic α-kleisin cohesin subunit Rad21/Scc1/Mcd1 by the meiotic paralog Rec8. In Drosophila, however, no clear Rec8 homolog has been identified so far. Therefore, we have analyzed the role of the mitotic Drosophila α-kleisin Rad21 during female meiosis. Inactivation of an engineered Rad21 variant by premature, ectopic cleavage during oogenesis results not only in loss of cohesin from meiotic chromatin, but also in precocious disassembly of the synaptonemal complex (SC). We demonstrate that the lateral SC component C(2)M can interact directly with Rad21, potentially explaining why Rad21 is required for SC maintenance. Intriguingly, the experimentally induced premature Rad21 elimination, as well as the expression of a Rad21 variant with destroyed separase consensus cleavage sites, do not interfere with chromosome segregation during meiosis, while successful mitotic divisions are completely prevented. Thus, chromatid cohesion during female meiosis does not depend on Rad21-containing cohesin.
复制后的姐妹染色单体从合成之时起就紧密相连,直至在下一次有丝分裂过程中分离。这种关联由环形的黏连蛋白复合体介导,该复合体似乎环绕着姐妹染色单体。在中期到后期转换时,黏连蛋白复合体的α-kleisin亚基被分离酶进行蛋白水解切割后,姐妹染色单体分离并被分配到子细胞核中。减数分裂过程中更为复杂的染色体分离被认为依赖于有丝分裂的α-kleisin黏连蛋白亚基Rad21/Scc1/Mcd1被减数分裂同源物Rec8所取代。然而,在果蝇中,到目前为止尚未鉴定出明确的Rec8同源物。因此,我们分析了有丝分裂的果蝇α-kleisin Rad21在雌性减数分裂中的作用。在卵子发生过程中,通过过早的异位切割使工程改造的Rad21变体失活,不仅导致减数分裂染色质上的黏连蛋白丢失,还导致联会复合体(SC)过早解体。我们证明,SC的侧向成分C(2)M可以直接与Rad21相互作用,这可能解释了为什么Rad21是维持SC所必需的。有趣的是,实验诱导的过早消除Rad21,以及表达具有被破坏的分离酶共有切割位点的Rad21变体,并不干扰减数分裂过程中的染色体分离,而有丝分裂的成功进行则被完全阻止。因此,雌性减数分裂过程中的染色单体黏连并不依赖于含Rad21的黏连蛋白。