Ursic D, Culbertson M R
Laboratory of Molecular Biology, University of Wisconsin, Madison 53706.
Mol Cell Biol. 1991 May;11(5):2629-40. doi: 10.1128/mcb.11.5.2629-2640.1991.
A Saccharomyces cerevisiae homolog to Drosophila melanogaster and mouse Tcp-1 encoding tailless complex polypeptide 1 (TCP1) has been identified, sequenced, and mapped. The mouse t complex has been under scrutiny for six decades because of its effects on embryogenesis and sperm differentiation and function. TCP1 is an essential gene in yeast cells and is located on chromosome 4R, linked to pet14. The TCP1-encoded proteins in yeast, Drosophila, and mouse cells share between 61 and 72% amino acid sequence identities, suggesting a primordial function for the TCP1 gene product. To assess function, we constructed a cold-impaired recessive mutation (tcp1-1) in the yeast gene. Cells carrying the tcp1-1 mutation grew linearly rather than exponentially at the restrictive temperature of 15 degrees C with a generation time of approximately 32 h in minimal medium. Both multinucleate and anucleate cells accumulated with time, suggesting that the linear growth kinetics may be explained by the generation of anucleate buds incapable of further cell division. In addition, the multinucleate and anucleate cells contained morphologically abnormal structures detected by anti-alpha-tubulin antibodies. The kinetics of appearance of these abnormalities suggest that they are a direct consequence of loss of function of the TCP1 protein and not a delayed, indirect consequence of cell death. We also observed that strains carrying tcp1-1 were hypersensitive to antimitotic compounds. Taken together, these observations imply that the TCP1 protein affects microtubule-mediated processes.
已鉴定、测序并定位了酿酒酵母中与黑腹果蝇和小鼠Tcp-1同源的基因,该基因编码无尾复合多肽1(TCP1)。由于小鼠t复合体对胚胎发生以及精子分化和功能的影响,六十年来一直受到密切关注。TCP1是酵母细胞中的必需基因,位于4R染色体上,与pet14连锁。酵母、果蝇和小鼠细胞中由TCP1编码的蛋白质的氨基酸序列同一性在61%至72%之间,这表明TCP1基因产物具有原始功能。为了评估其功能,我们在酵母基因中构建了一个冷敏感隐性突变(tcp1-1)。携带tcp1-1突变的细胞在15摄氏度的限制温度下以线性而非指数方式生长,在基本培养基中的代时约为32小时。随着时间的推移,多核细胞和无核细胞都有所积累,这表明线性生长动力学可能是由无法进一步细胞分裂的无核芽的产生所解释的。此外,多核细胞和无核细胞含有用抗α-微管蛋白抗体检测到的形态异常结构。这些异常出现的动力学表明,它们是TCP1蛋白功能丧失的直接后果,而不是细胞死亡的延迟间接后果。我们还观察到携带tcp1-1的菌株对抗有丝分裂化合物高度敏感。综上所述,这些观察结果表明TCP1蛋白影响微管介导的过程。