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首页> 外文期刊>Molecular and Cellular Biology >The Cyc8 (Ssn6)-Tup1 corepressor complex is composed of one Cyc8 and four Tup1 subunits.
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The Cyc8 (Ssn6)-Tup1 corepressor complex is composed of one Cyc8 and four Tup1 subunits.

机译：Cyc8（Ssn6）-Tup1核心复合体由一个Cyc8和四个Tup1亚基组成。

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摘要

The Cyc8 (Ssn6)-Tup1 corepressor complex is required for repression in several important regulatory systems in yeast cells, including glucose repression and mating type. Cyc8-Tup1 is recruited to target genes by interaction with diverse repressor proteins that bind directly to DNA. Since the complex has a large apparent molecular mass of 1,200 kDa on nondenaturing gels (F. E. Williams, U. Varanasi, and R. J. Trumbly, Mol. Cell. Biol. 11:3307-3316, 1991), we used a variety of approaches to determine its actual subunit composition. Immunoprecipitation of epitope-tagged complex and reconstitution of the complex from in vitro-translated proteins demonstrated that only the Cyc8 and Tup1 proteins were present in the complex. Hydrodynamic properties showed that these proteins have unusually large Stokes radii, low sedimentation coefficients, and high frictional ratios, all characteristic of asymmetry which partly accounts for the apparent high molecular weight. Calculation of native molecular weights from these properties indicated that the Cyc8-Tup1 complex is composed of one Cyc8 subunit and four Tup1 subunits. This composition was confirmed by reconstitution of the complex from Cyc8 and Tup1 expressed in vitro and analysis by one- and two-dimensional gel electrophoresis.
机译：Cyc8（Ssn6）-Tup1核心复合物是酵母细胞中几个重要调控系统（包括葡萄糖阻遏和交配类型）阻遏所必需的。 Cyc8-Tup1通过与直接与DNA结合的多种阻遏蛋白相互作用而募集到目标基因。由于该复合物在非变性凝胶上具有1200 kDa的大表观分子量（FE Williams，U。Varanasi和RJ Trumbly，Mol。Cell。Biol。11：3307-3316，1991），因此我们使用了多种方法来确定其实际的亚基组成。抗原决定簇标记的复合物的免疫沉淀和复合物从体外翻译蛋白的重建表明，复合物中仅存在Cyc8和Tup1蛋白。流体力学性质表明，这些蛋白质具有异常大的斯托克斯半径，低沉降系数和高摩擦比，所有这些都是不对称的特征，部分原因是表观的高分子量。从这些性质计算天然分子量表明，Cyc8-Tup1复合物由一个Cyc8亚基和四个Tup1亚基组成。通过重组体外表达的Cyc8和Tup1的复合物并通过一维和二维凝胶电泳进行分析，证实了该组成。

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