Su S S, Mitchell A P
Institute of Cancer Research, College of Physicians and Surgeons, Columbia University, New York, NY 10032.
Nucleic Acids Res. 1993 Aug 11;21(16):3789-97. doi: 10.1093/nar/21.16.3789.
In the yeast Saccharomyces cerevisiae, genetic studies suggest that the RIM1 gene encodes a positive regulator of meiosis. rim1 mutations cause reduced expression of IME1, which is required for expression of many meiotic genes, and thus lead to a partial defect in meiosis and spore formation. We report the sequence of RIM1 and functional analysis of its coding region. The RIM1 gene product (RIM1) contains three regions similar to C2H2 zinc fingers. Serine substitutions for cysteine in each of the putative zinc fingers abolish RIM1 function. The carboxyl-terminus of RIM1 is enriched in acidic amino acids and is required for full RIM1 activity. RIM1 also contains two putative cAMP-dependent protein kinase (cAPK) phosphorylation sites. At one site, substitution of alanine for serine does not affect RIM1 activity; at the other site, this substitution impairs activity. This analysis of RIM1 suggests that the protein may function as a transcriptional activator. We have used the cloned RIM1 gene to create a complete rim1 deletion. This null allele, like previously isolated rim1 mutations, causes a partial meiotic defect. In addition to RIM1, maximum IME1 expression requires the MCK1 and IME4 gene products. Defects associated with rim1, mck1, and ime4 mutations in expression of a meiotic reporter gene (ime2-lacZ) and in sporulation are additive. These findings suggest that RIM1 acts independently of MCK1 and IME4 to stimulate IME1 expression.
在酿酒酵母中,遗传学研究表明RIM1基因编码减数分裂的一个正向调节因子。rim1突变导致IME1表达降低,而IME1是许多减数分裂基因表达所必需的,因此导致减数分裂和孢子形成出现部分缺陷。我们报道了RIM1的序列及其编码区的功能分析。RIM1基因产物(RIM1)包含三个与C2H2锌指相似的区域。在每个假定的锌指中,用丝氨酸取代半胱氨酸会消除RIM1的功能。RIM1的羧基末端富含酸性氨基酸,是RIM1完全活性所必需的。RIM1还包含两个假定的依赖于cAMP的蛋白激酶(cAPK)磷酸化位点。在一个位点,用丙氨酸取代丝氨酸不影响RIM1活性;在另一个位点,这种取代会损害活性。对RIM1的这种分析表明,该蛋白可能作为转录激活因子发挥作用。我们已使用克隆的RIM1基因创建了一个完整的rim1缺失。这个无效等位基因,与先前分离的rim1突变一样,会导致部分减数分裂缺陷。除了RIM1外,IME1的最大表达还需要MCK1和IME4基因产物。与rim1、mck1和ime4突变相关的减数分裂报告基因(ime2-lacZ)表达缺陷和孢子形成缺陷是累加的。这些发现表明,RIM1独立于MCK1和IME4发挥作用以刺激IME1表达。