Brown M T, Goetsch L, Hartwell L H
Department of Genetics, University of Washington, Seattle 98195.
J Cell Biol. 1993 Oct;123(2):387-403. doi: 10.1083/jcb.123.2.387.
The function of the essential MIF2 gene in the Saccharomyces cerevisiae cell cycle was examined by overepressing or creating a deficit of MIF2 gene product. When MIF2 was overexpressed, chromosomes missegregated during mitosis and cells accumulated in the G2 and M phases of the cell cycle. Temperature sensitive mutants isolated by in vitro mutagenesis delayed cell cycle progression when grown at the restrictive temperature, accumulated as large budded cells that had completed DNA replication but not chromosome segregation, and lost viability as they passed through mitosis. Mutant cells also showed increased levels of mitotic chromosome loss, supersensitivity to the microtubule destabilizing drug MBC, and morphologically aberrant spindles. mif2 mutant spindles arrested development immediately before anaphase spindle elongation, and then frequently broke apart into two disconnected short half spindles with misoriented spindle pole bodies. These findings indicate that MIF2 is required for structural integrity of the spindle during anaphase spindle elongation. The deduced Mif2 protein sequence shared no extensive homologies with previously identified proteins but did contain a short region of homology to a motif involved in binding AT rich DNA by the Drosophila D1 and mammalian HMGI chromosomal proteins.
通过过表达或造成MIF2基因产物缺陷,研究了酿酒酵母细胞周期中必需的MIF2基因的功能。当MIF2过表达时,有丝分裂期间染色体发生错分,细胞在细胞周期的G2期和M期积累。通过体外诱变分离的温度敏感突变体在限制温度下生长时延迟了细胞周期进程,积累为已完成DNA复制但未进行染色体分离的大芽殖细胞,并在通过有丝分裂时丧失活力。突变细胞还表现出有丝分裂染色体丢失水平增加、对微管去稳定药物MBC超敏感以及纺锤体形态异常。mif2突变体纺锤体在后期纺锤体伸长前立即停止发育,然后经常分裂成两个断开的短半纺锤体,纺锤极体方向错误。这些发现表明,在后期纺锤体伸长过程中,MIF2是纺锤体结构完整性所必需的。推导的Mif2蛋白序列与先前鉴定的蛋白没有广泛的同源性,但确实包含一个与果蝇D1和哺乳动物HMGI染色体蛋白结合富含AT的DNA的基序有短同源区域。