Osmani A H, Osmani S A, Morris N R
Department of Pharmacology, Robert Wood Johnson Medical School, University of Medicine and Dentistry of New Jersey, Piscataway 08854.
J Cell Biol. 1990 Aug;111(2):543-51. doi: 10.1083/jcb.111.2.543.
A temperature-sensitive mutation in the nudC gene (nudC3) of Aspergillus nidulans specifically prevents the microtubule-based movement of nuclei in this organism at the restrictive temperature. The mutation does not affect short term growth, nuclear division, or the movement of other subcellular organelles. Immunofluorescence analysis of cells blocked at the restrictive temperature, using antitubulin antibodies, shows that the inability of nuclei to move under these conditions is not related to an inability of a particular class of microtubule to form. The inability to move nuclei in this mutant is also shown to be independent of both mitosis and the number of nuclei in the cell as a double mutant carrying both nudC3 and a cell cycle-specific mutation blocks with a single immotile nucleus at the restrictive temperature. The molecular cloning of the nudC gene and sequence analysis reveal that it encodes a previously unidentified protein of 22 kd. Affinity-purified antisera reactive to the nudC protein cross reacts to a single protein of 22 kD in Aspergillus protein extracts. This purified sera failed to reveal a subcellular location for the nudC protein at the level of indirect immunofluorescence. The data presented suggest that the 22-kD nudC gene product functions by interacting between microtubules and nuclei and/or is involved in the generation of force used to move nuclei during interphase.
构巢曲霉nudC基因(nudC3)中的一个温度敏感突变,在限制温度下特异性地阻止了该生物体中基于微管的细胞核移动。该突变不影响短期生长、核分裂或其他亚细胞细胞器的移动。使用抗微管蛋白抗体对在限制温度下受阻的细胞进行免疫荧光分析表明,在这些条件下细胞核无法移动与某一类微管无法形成无关。在该突变体中细胞核无法移动也被证明与有丝分裂和细胞中的细胞核数量均无关,因为携带nudC3和细胞周期特异性突变的双突变体在限制温度下会因单个不能移动的细胞核而受阻。nudC基因的分子克隆和序列分析表明,它编码一种先前未鉴定的22kd蛋白质。与nudC蛋白反应的亲和纯化抗血清与构巢曲霉蛋白提取物中的一种22kD的单一蛋白质发生交叉反应。这种纯化的血清在间接免疫荧光水平上未能揭示nudC蛋白的亚细胞定位。所呈现的数据表明,22-kD nudC基因产物通过在微管和细胞核之间相互作用发挥功能,和/或参与在间期用于移动细胞核的力的产生。