Ali Emine I, Loidl Josef, Howard-Till Rachel A
Department of Chromosome Biology, Vienna Biocenter, University of Vienna, Vienna, Austria.
Chromosoma. 2018 Dec;127(4):421-435. doi: 10.1007/s00412-018-0673-x. Epub 2018 Jun 12.
In order to understand its diverse functions, we have studied cohesin in the evolutionarily distant ciliate model organism Tetrahymena thermophila. In this binucleate cell, the heritable germline genome is maintained separately from the transcriptionally active somatic genome. In a previous study, we showed that a minimal cohesin complex in Tetrahymena consisted of homologs of Smc1, Smc3, and Rec8, which are present only in the germline nucleus, where they are needed for normal chromosome segregation as well as meiotic DNA repair. In this study, we confirm that a putative homolog of Scc3 is a member of this complex. In the absence of Scc3, Smc1 and Rec8 fail to localize to germline nuclei, Rec8 is hypo-phosphorylated, and cells show phenotypes similar to depletion of Smc1 and Rec8. We also identify a homolog of Scc2, which in other organisms is part of a heterodimeric complex (Scc2/Scc4) that helps load cohesin onto chromatin. In Tetrahymena, Scc2 interacts with Rec8 and Scc3, and its absence causes defects in mitotic and meiotic divisions. Scc2 is not required for chromosomal association of cohesin, but Rec8 is hypo-phosphorylated in its absence. Moreover, we did not identify a homolog of the cohesin loader Scc4, and no evidence was found of auxiliary factors, such as Eco1, Pds5, or WAPL. We propose that in Tetrahymena, a single, minimal cohesin complex performs all necessary functions for germline mitosis and meiosis, but is dispensable for transcription regulation and chromatin organization of the somatic genome.
为了了解黏连蛋白的多种功能,我们在进化上距离较远的纤毛虫模式生物嗜热四膜虫中研究了黏连蛋白。在这种双核细胞中,可遗传的种系基因组与转录活跃的体细胞基因组是分开维持的。在之前的一项研究中,我们表明四膜虫中的最小黏连蛋白复合体由Smc1、Smc3和Rec8的同源物组成,这些同源物仅存在于种系细胞核中,在那里它们是正常染色体分离以及减数分裂DNA修复所必需的。在本研究中,我们证实Scc3的一个推定同源物是该复合体的成员。在没有Scc3的情况下,Smc1和Rec8无法定位于种系细胞核,Rec8磷酸化不足,并且细胞表现出与Smc1和Rec8缺失相似的表型。我们还鉴定出了Scc2的一个同源物,在其他生物体中,Scc2是异二聚体复合体(Scc2/Scc4)的一部分,该复合体有助于将黏连蛋白加载到染色质上。在四膜虫中,Scc2与Rec8和Scc3相互作用,其缺失会导致有丝分裂和减数分裂出现缺陷。黏连蛋白与染色体的结合不需要Scc2,但在其缺失时Rec8磷酸化不足。此外,我们没有鉴定出黏连蛋白加载因子Scc4的同源物,也没有发现诸如Eco1、Pds5或WAPL等辅助因子的证据。我们提出,在四膜虫中,一个单一的最小黏连蛋白复合体执行种系有丝分裂和减数分裂的所有必要功能,但对于体细胞基因组的转录调控和染色质组织是可有可无的。