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首页> 外文期刊>Proceedings of the National Academy of Sciences of the United States of America >Probing the Saccharomyces cerevisiae centromeric DNA (CEM DNA)-binding factor 3 (CBF3) kinetochore complex by using atomic force microscopy
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Probing the Saccharomyces cerevisiae centromeric DNA (CEM DNA)-binding factor 3 (CBF3) kinetochore complex by using atomic force microscopy

机译：用原子力显微镜探查酿酒酵母着丝粒DNA（CEM DNA）结合因子3（CBF3）线粒体。

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摘要

Yeast centromeric DNA (CEN DNA) binding factor 3 (CBF3) is a multisubunit protein complex that binds to the essential CDElll element in CEN DNA. The four CBF3 proteins are required for accurate chromosome segregation and are considered to be core components of the yeast kinetochore. We have examined the structure of the CBF3CEN DNA complex by atomic force microscopy. Assembly of CBF3-CEN DNA complexes was Performed by combining purified CBF3 proteins with a DNA fragment that includes the CEN region from yeast chromosome Ill. Atomic force microscopy images showed DNA molecules with attached globular bodies. The contour length of the DNA containing the complex is approximately equals 9 shorter than the DNA alone, suggesting some winding of DNA within the complex. The measured location of the Single binding site indicates that the complex is located asymmetrically to the right of CDElll extending away from CDEI and CDEll, which is consistent with previous data. The CEN DNA is bent approximately equals 55 degrees at the site of complex formation. A significant fraction of the complexes are linked in pairs, showing three to four DNA arms, with molecular volumes approximately three times the mean volumes of two- armed complexes. These multi-armed complexes indicate that CBF3 can bind two DNA molecules together in vitro and, thus, may be involved in holding together chromatid pairs during mitosis.
机译：酵母着丝粒DNA（CEN DNA）结合因子3（CBF3）是一种多亚基蛋白质复合物，可与CEN DNA中的基本CDEll1元件结合。四种CBF3蛋白是准确染色体分离所必需的，被认为是酵母动线粒的核心成分。我们已经通过原子力显微镜检查了CBF3CEN DNA复合物的结构。 CBF3-CEN DNA复合物的组装是通过将纯化的CBF3蛋白与一个包含来自酵母染色体Ill的CEN区域的DNA片段相结合而完成的。含有复合物的DNA的轮廓长度比单独的DNA短约9倍，这表明DNA在复合物中有些缠绕。单一结合位点的测量位置表明该复合物不对称地位于CDEll1的右侧，远离CDEI和CDEll，与以前的数据一致。 CEN DNA在复合物形成部位弯曲大约等于55度。大部分复合物成对连接，显示三到四个DNA臂，分子体积约为两臂复合物平均体积的三倍。这些多臂复合物表明CBF3可以在体外将两个DNA分子结合在一起，因此可能参与有丝分裂期间将染色单体对结合在一起。

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