Crystal Structure of the Cohesin Gatekeeper Pds5 and in Complex with Kleisin Scc1.
Crystal Structure of the Cohesin Gatekeeper Pds5 and in Complex with Kleisin Scc1.
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DOI：
10.1016/j.celrep.2016.02.020
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发表时间：
2016-03-08
期刊：
影响因子：
8.8
通讯作者：
Löwe J
中科院分区：
文献类型：
--
作者：
Lee BG;Roig MB;Jansma M;Petela N;Metson J;Nasmyth K;Löwe J
姐妹染色单体内聚是由内聚蛋白介导的，其Smc1、Smc3和柔素（Scc1）亚基形成一个环结构，可以捕获姐妹dna。这个环要么是由分离酶打开的，分离酶在后期分裂Scc1，要么是由涉及Wapl、Scc3和Pds5的释放活动打开的，这些释放活动结合Scc1并打开Scc1与Smc3的接口。我们展示了来自酵母L.耐热菌的Pds5在存在和不存在与Pds5相互作用的保守Scc1区域的晶体结构。Scc1沿着Pds5 HEAT重复折叠的脊柱结合，并楔入Pds5的脊柱和c端钩之间。我们分离的突变体证实了观察到的Scc1结合模式，并通过免疫沉淀和校准的ChIP-seq验证了它们对内聚素的影响。Pds5的结构也显示出与Scc3的结构相似性，Scc3是内聚蛋白的另一个大的HEAT重复蛋白，很可能与Scc2相似。通过coIP和ChIP-seq对Pds5和Scc1的结构突变体进行了分析，发现Pds5与Scc3具有一定的相似性，另一个较大的HEAT重复黏结蛋白亚基cohesin在有丝分裂和减数分裂过程中以环状形式包裹DNA，促进姐妹染色单体的黏结。Lee等人描述了内聚蛋白亚基Pds5与Scc1复合物的结构，内聚蛋白的收缩蛋白，破坏Pds5:Scc1相互作用的突变，以及与另一个内聚蛋白亚基Scc3的结构相似性。
Sister chromatid cohesion is mediated by cohesin, whose Smc1, Smc3, and kleisin (Scc1) subunits form a ring structure that entraps sister DNAs. The ring is opened either by separase, which cleaves Scc1 during anaphase, or by a releasing activity involving Wapl, Scc3, and Pds5, which bind to Scc1 and open its interface with Smc3. We present crystal structures of Pds5 from the yeast L. thermotolerans in the presence and absence of the conserved Scc1 region that interacts with Pds5. Scc1 binds along the spine of the Pds5 HEAT repeat fold and is wedged between the spine and C-terminal hook of Pds5. We have isolated mutants that confirm the observed binding mode of Scc1 and verified their effect on cohesin by immunoprecipitation and calibrated ChIP-seq. The Pds5 structure also reveals architectural similarities to Scc3, the other large HEAT repeat protein of cohesin and, most likely, Scc2. The crystal structure of the cohesin subunit Pds5 was determined The crystal structure of Pds5 in complex with Scc1 binding region was determined Structure-based mutants in Pds5 and Scc1 were analyzed by coIP and ChIP-seq Pds5 shows some similarity to Scc3, the other large HEAT repeat cohesin subunit Cohesin entraps DNA in a ring, facilitating sister chromatid cohesion during mitosis and meiosis. Lee et al. describe the structure of the cohesin subunit Pds5 in complex with Scc1, cohesin’s kleisin, mutants disrupting the Pds5:Scc1 interaction, and structural similarity to another cohesin subunit, Scc3.