McMillan J N, Tatchell K
Department of Microbiology, North Carolina State University, Raleigh 27695.
J Cell Biol. 1994 Apr;125(1):143-58. doi: 10.1083/jcb.125.1.143.
JNM1, a novel gene on chromosome XIII in the yeast Saccharomyces cerevisiae, is required for proper nuclear migration. jnm1 null mutants have a temperature-dependent defect in nuclear migration and an accompanying alteration in astral microtubules. At 30 degrees C, a significant proportion of the mitotic spindles is not properly located at the neck between the mother cell and the bud. This defect is more severe at low temperature. At 11 degrees C, 60% of the cells accumulate with large buds, most of which have two DAPI staining regions in the mother cell. Although mitosis is delayed and nuclear migration is defective in jnm1 mutant, we rarely observe more than two nuclei in a cell, nor do we frequently observe anuclear cells. No loss of viability is observed at 11 degrees C and cells continue to grow exponentially with increased doubling time. At low temperature the large budded cells of jnm1 mutants exhibit extremely long astral microtubules that often wind around the periphery of the cell. jnm1 mutants are not defective in chromosome segregation during mitosis, as assayed by the rate of chromosome loss, or nuclear migration during conjugation, as assayed by the rate of mating and cytoduction. The phenotype of a jnm1 mutant is strikingly similar to that for mutants in the dynein heavy chain gene (Eshel, D., L. A. Urrestarazu, S. Vissers, J.-C. Jauniaux, J. C. van Vliet-Reedijk, R. J. Plants, and I. R. Gibbons. 1993. Proc. Natl. Acad. Sci. USA. 90:11172-11176; Li, Y. Y., E. Yeh, T. Hays, and K. Bloom. 1993. Proc. Natl. Acad. Sci. USA. 90:10096-10100). The JNM1 gene product is predicted to encode a 44-kD protein containing three coiled coil domains. A JNM1:lacZ gene fusion is able to complement the cold sensitivity and microtubule phenotype of a jnm1 deletion strain. This hybrid protein localizes to a single spot in the cell, most often near the spindle pole body in unbudded cells and in the bud in large budded cells. Together these results point to a specific role for Jnm1p in spindle migration, possibly as a subunit or accessory protein for yeast dynein.
JNM1是酿酒酵母(Saccharomyces cerevisiae)XIII号染色体上的一个新基因,是正常核迁移所必需的。jnm1缺失突变体在核迁移方面存在温度依赖性缺陷,且星体微管也有相应改变。在30℃时,相当一部分有丝分裂纺锤体未正确定位在母细胞与芽之间的颈部。在低温下这种缺陷更严重。在11℃时,60%的细胞积累形成大芽,其中大多数母细胞中有两个DAPI染色区域。尽管jnm1突变体中细胞分裂延迟且核迁移有缺陷,但我们很少在一个细胞中观察到超过两个细胞核,也不常观察到无核细胞。在11℃时未观察到活力丧失,细胞继续以增加的倍增时间指数生长。在低温下,jnm1突变体的大芽细胞表现出极长的星体微管,这些微管常常缠绕在细胞周边。通过染色体丢失率检测,jnm1突变体在有丝分裂期间的染色体分离无缺陷;通过交配率和细胞质融合率检测,其在接合过程中的核迁移也无缺陷。jnm1突变体的表型与动力蛋白重链基因突变体的表型惊人地相似(埃舍尔,D.,L.A.乌雷斯塔拉祖,S.维瑟斯,J.-C.若尼奥,J.C.范弗利特 - 里德伊克,R.J.普兰茨,和I.R.吉本斯。1993年。美国国家科学院院刊。90:11172 - 11176;李,Y.Y.,E.叶,T.海斯,和K.布鲁姆。1993年。美国国家科学院院刊。90:10096 - 10100)。预测JNM1基因产物编码一个含有三个卷曲螺旋结构域的44-kD蛋白。一个JNM1:lacZ基因融合体能够弥补jnm1缺失菌株的冷敏感性和微管表型。这种杂交蛋白定位于细胞中的一个单点,在未出芽细胞中最常靠近纺锤极体,在大芽细胞中位于芽内。这些结果共同表明Jnm1p在纺锤体迁移中具有特定作用,可能作为酵母动力蛋白的一个亚基或辅助蛋白。