Doe C L, Wang G, Chow C, Fricker M D, Singh P B, Mellor E J
Microbiology Unit, Department of Biochemistry, University of Oxford, South Parks Road, Oxford OX1 3QU, UK, Department of Development and Genetics, The Babraham Institute, Babraham Hall, Babraham, Cambridge CB2 4AT, UK.
Nucleic Acids Res. 1998 Sep 15;26(18):4222-9. doi: 10.1093/nar/26.18.4222.
In eukaryotes, the segregation of chromosomes is co-ordinated by the centromere and must proceed accurately if aneuploidy and cell death are to be avoided. The fission yeast centromere is complex, containing highly repetitive regions of DNA showing the characteristics of heterochromatin. Two proteins, Swi6p and Clr4p, that are associated with the fission yeast centromere also contain a chromo (chromatin organisation modifier) domain and are required for centromere function. We have analysed a novel fission yeast gene encoding a putative chromo domain called chp 1(+) (chromo domain protein in Schizosaccharomyces p ombe ). In the absence of Chp1p protein, cells are viable but show chromosome segregation defects such as lagging chromosomes on the spindle during anaphase and high rates of minichromosome loss, phenotypes which are also displayed by swi 6 and clr 4. A fusion protein between green fluorescent protein (GFP) and Chp1p, like Swi6p, is localized to discrete sites within the nucleus. In contrast to Swi6p and Clr4p, Chp1p is not required to repress silent mating-type genes. We demonstrate a genetic interaction between chp 1(+) and alpha-tubulin ( nda 2(+)) and between swi 6(+) and beta-tubulin ( nda 3(+)). Chp1p and Swi6p proteins may be components of the kinetochore which captures and stabilizes the microtubules of the spindle.
在真核生物中,染色体的分离由着丝粒协调,若要避免非整倍体和细胞死亡,这一过程必须精确进行。裂殖酵母的着丝粒很复杂,包含具有异染色质特征的高度重复DNA区域。与裂殖酵母着丝粒相关的两种蛋白质Swi6p和Clr4p也含有一个染色质组织修饰(chromo)结构域,并且是着丝粒功能所必需的。我们分析了一个新的裂殖酵母基因,它编码一种假定的名为chp1⁺(裂殖酵母中的染色质结构域蛋白)的染色质结构域。在缺乏Chp1p蛋白的情况下,细胞能够存活,但表现出染色体分离缺陷,如后期纺锤体上的滞后染色体和高频率的小染色体丢失,这些表型也出现在swi6和clr4突变体中。绿色荧光蛋白(GFP)与Chp1p的融合蛋白,与Swi6p一样,定位于细胞核内的离散位点。与Swi6p和Clr4p不同,Chp1p不是抑制沉默交配型基因所必需的。我们证明了chp1⁺与α-微管蛋白(nda2⁺)以及swi6⁺与β-微管蛋白(nda3⁺)之间存在遗传相互作用。Chp1p和Swi6p蛋白可能是动粒的组成成分,动粒负责捕获并稳定纺锤体的微管。