Meluh P B, Yang P, Glowczewski L, Koshland D, Smith M M
Howard Hughes Medical Institute, Carnegie Institution of Washington, Department of Embryology, Baltimore, Maryland 21210, USA.
Cell. 1998 Sep 4;94(5):607-13. doi: 10.1016/s0092-8674(00)81602-5.
Histones are fundamental structural components of chromatin and are expected to play important roles in chromosome dynamics. Here, we present direct evidence that Cse4p, a histone H3 variant, is a structural component of the core centromere of S. cerevisiae. In histone H4 and Cse4p mutants, the core centromere chromatin structure is disrupted at restrictive temperature. Overexpression of Cse4p suppresses this defect in the H4 mutant, implying that the two proteins act together in centromere structure. We show by chromatin immunoprecipitation experiments that Cse4p is specifically cross-linked to centromeric DNA. Furthermore, by immunofluorescence microscopy, Cse4p is found in discrete foci consistent with that expected for centromeres. These results suggest the kinetochore is assembled on a specialized centromeric nucleosome containing Cse4p.
组蛋白是染色质的基本结构成分,有望在染色体动态变化中发挥重要作用。在此,我们提供直接证据表明,组蛋白H3变体Cse4p是酿酒酵母核心着丝粒的结构成分。在组蛋白H4和Cse4p突变体中,核心着丝粒染色质结构在限制温度下被破坏。Cse4p的过表达抑制了H4突变体中的这一缺陷,这意味着这两种蛋白质在着丝粒结构中共同发挥作用。我们通过染色质免疫沉淀实验表明,Cse4p与着丝粒DNA特异性交联。此外,通过免疫荧光显微镜观察,发现Cse4p存在于与着丝粒预期相符的离散斑点中。这些结果表明,动粒是在含有Cse4p的特殊着丝粒核小体上组装的。