有丝分裂酵母着丝粒特异性核小体的生物物理特性分析。
Biophysical characterization of the centromere-specific nucleosome from budding yeast.
机构信息
Macromolecular Structure and Function Laboratory, Cancer Research UK London Research Institute, 44 Lincoln's Inn Fields, London WC2A 3LY, United Kingdom.
出版信息
J Biol Chem. 2011 Feb 4;286(5):4021-6. doi: 10.1074/jbc.M110.189340. Epub 2010 Nov 29.
Abstract
The centromeric DNA of all eukaryotes is assembled upon a specialized nucleosome containing a histone H3 variant known as CenH3. Despite the importance and conserved nature of this protein, the characteristics of the centromeric nucleosome are still poorly understood. In particular, the stoichiometry and DNA-binding properties of the CenH3 nucleosome have been the subject of some debate. We have characterized the budding yeast centromeric nucleosome by biochemical and biophysical methods and show that it forms a stable octamer containing two copies of the Cse4 protein and wraps DNA in a left-handed supercoil, similar to the canonical H3 nucleosome. The DNA-binding properties of the recombinant nucleosome are identical to those observed in vivo demonstrating that the octameric structure is physiologically relevant.
摘要
所有真核生物的着丝粒 DNA 都组装在含有一种组蛋白 H3 变体的特殊核小体上，这种变体被称为 CenH3。尽管这种蛋白质非常重要且具有保守性，但着丝粒核小体的特征仍知之甚少。特别是，CenH3 核小体的计量学和 DNA 结合特性一直存在争议。我们使用生化和生物物理方法对芽殖酵母着丝粒核小体进行了表征，结果表明它形成了一个稳定的八聚体，包含两个 Cse4 蛋白拷贝，并以左手超螺旋的形式包裹 DNA，类似于典型的 H3 核小体。重组核小体的 DNA 结合特性与体内观察到的特性相同，这表明八聚体结构具有生理相关性。
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