Shero J H, Hieter P
Department of Molecular Biology and Genetics, Johns Hopkins University School of Medicine, Baltimore, Maryland 21205.
Genes Dev. 1991 Apr;5(4):549-60. doi: 10.1101/gad.5.4.549.
A new approach to identify genes involved in Saccharomyces cerevisiae kinetochore function is discussed. A genetic screen was designed to recover extragenic dosage suppressors of a CEN DNA mutation. This method identified two suppressors, designated MCK1 and CMS2. Increased dosage of MCK1 specifically suppressed two similar CEN DNA mutations in CDEIII, but not comparably defective CEN DNA mutations in CDEI or CDEII. A strain containing a null allele of MCK1 was viable under standard growth conditions, had a cold-sensitive phenotype (conditional lethality at 11 degrees C), and grew slowly on Benomyl (a microtubule-destabilizing drug). Furthermore, when grown at 18 degrees C or in the presence of Benomyl, the null mutant exhibited a dramatic increase in the rate of mitotic chromosome loss. The allele-specific suppression and chromosome instability phenotypes suggest that MCK1 plays a role in mitotic chromosome segregation specific to CDEIII function. The MCK1 gene encodes a putative protein-serine/threonine kinase, which suggests a possible role for the MCK1 protein in regulating the activity of centromere-binding proteins by phosphorylation. MCK1 was identified and cloned independently for its involvement in the induction of meiosis and is identical to a gene that encodes a phosphotyrosyl protein with protein kinase activity.
本文讨论了一种鉴定参与酿酒酵母动粒功能基因的新方法。设计了一个遗传筛选来寻找CEN DNA突变的基因外剂量抑制子。该方法鉴定出两个抑制子,命名为MCK1和CMS2。增加MCK1的剂量可特异性抑制CDEIII中两个相似的CEN DNA突变,但不能抑制CDEI或CDEII中存在类似缺陷的CEN DNA突变。含有MCK1无效等位基因的菌株在标准生长条件下可存活,具有冷敏感表型(在11摄氏度时条件致死),并且在苯菌灵(一种微管破坏药物)上生长缓慢。此外,当在18摄氏度或存在苯菌灵的条件下生长时,无效突变体的有丝分裂染色体丢失率显著增加。等位基因特异性抑制和染色体不稳定性表型表明,MCK1在特定于CDEIII功能的有丝分裂染色体分离中起作用。MCK1基因编码一种假定的蛋白丝氨酸/苏氨酸激酶,这表明MCK1蛋白可能通过磷酸化作用调节着丝粒结合蛋白的活性。MCK1因其参与减数分裂的诱导而被独立鉴定和克隆,并且与一个编码具有蛋白激酶活性的磷酸酪氨酸蛋白的基因相同。