Liu Zhe, Makaroff Christopher A
Department of Chemistry and Biochemistry, Miami University, Oxford, Ohio 45056, USA.
Plant Cell. 2006 May;18(5):1213-25. doi: 10.1105/tpc.105.036913. Epub 2006 Mar 31.
To investigate how and when sister chromatid cohesion is released from chromosomes in plants, we isolated the Arabidopsis thaliana homolog of separase (AESP) and investigated its role in somatic and meiotic cells. AESP is similar to separase proteins identified in other organisms but contains several additional structural motifs. The characterization of two Arabidopsis T-DNA insertion alleles for AESP demonstrated that it is an essential gene. Seeds homozygous for T-DNA insertions in AESP exhibited embryo arrest at the globular stage. The endosperm also exhibited a weak titan-like phenotype. Transgenic plants expressing AESP RNA interference (RNAi) from the meiosis-specific DMC1 promoter exhibited alterations in chromosome segregation during meiosis I and II that resulted in polyads containing from one to eight microspores. Consistent with its predicted role in the release of sister chromatid cohesion, immunolocalization studies showed that the removal of SYN1 from chromosome arms and the centromeres is inhibited in the RNAi mutants. However, the release of SYN1 during diplotene occurred normally, indicating that this process is independent of AESP. Therefore, our results demonstrate that AESP plays an essential role in embryo development and provide direct evidence that AESP is required for the removal of cohesin from meiotic chromosomes.
为了研究植物中姐妹染色单体黏连是如何以及何时从染色体上释放的,我们分离了拟南芥中分离酶的同源物(AESP),并研究了它在体细胞和减数分裂细胞中的作用。AESP与在其他生物体中鉴定出的分离酶蛋白相似,但含有几个额外的结构基序。对AESP的两个拟南芥T-DNA插入等位基因的表征表明它是一个必需基因。AESP中T-DNA插入纯合的种子在球形期表现出胚胎停滞。胚乳也表现出一种较弱的类泰坦型表型。从减数分裂特异性DMC1启动子表达AESP RNA干扰(RNAi)的转基因植物在减数分裂I和II期间染色体分离出现改变,导致形成含有一到八个小孢子的多联体。与其在姐妹染色单体黏连释放中的预测作用一致,免疫定位研究表明RNAi突变体中染色体臂和着丝粒上SYN1的去除受到抑制。然而,双线期期间SYN1的释放正常发生,表明这个过程独立于AESP。因此,我们的结果表明AESP在胚胎发育中起重要作用,并提供了直接证据表明AESP是减数分裂染色体上黏连蛋白去除所必需的。