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A streamlined cohesin apparatus is sufficient for mitosis and meiosis in the protist Tetrahymena

机译:流线型粘附素装置足以应对四面膜虫的有丝分裂和减数分裂

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摘要

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.Electronic supplementary materialThe online version of this article (10.1007/s00412-018-0673-x) contains supplementary material, which is available to authorized users.
机译:为了了解其多种功能,我们在进化距离较远的纤毛模型生物嗜热四膜虫中研究了粘着蛋白。在该双核细胞中,可遗传的种系基因组与转录活性的体细胞基因组分开维护。在先前的研究中,我们表明四膜虫中的最小黏附素复合物由Smc1,Smc3和Rec8的同源物组成,它们仅存在于种系核中,而正常染色体分离和减数分裂DNA修复则需要它们。在这项研究中,我们确认推定的Scc3同源物是此复合物的成员。在缺少Scc3的情况下,Smc1和Rec8无法定位到种系核,Rec8被磷酸化,并且细胞表现出类似于Smc1和Rec8耗竭的表型。我们还确定了Scc2的同系物,它在其他生物中是异二聚体复合物(Scc2 / Scc4)的一部分,该异二聚体有助于将黏着素加载到染色质上。在四膜虫中,Scc2与Rec8和Scc3相互作用,并且其缺失会导致有丝分裂和减数分裂分裂的缺陷。 Scc2对于黏附蛋白的染色体缔合不是必需的,但是Rec8在不存在的情况下被磷酸化。此外,我们没有鉴定黏附素装载剂Scc4的同源物,也没有发现辅助因子如Eco1,Pds5或WAPL的证据。我们建议在四膜虫中,单一的最小黏附素复合物执行种系有丝分裂和减数分裂的所有必需功能,但对于体细胞基因组的转录调控和染色质组织是必不可少的。电子补充材料本文的在线版本(10.1007 / s00412-018 -0673-x)包含补充材料,授权用户可以使用。

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