Grandin N, Charbonneau M
Yeast Cell Cycle Group, UMR CNRS/ENS No. 8510, Ecole Normale Supérieure de Lyon, France.
Mol Gen Genet. 1999 Mar;261(2):402-7. doi: 10.1007/s004380050981.
Dbf2, a cell cycle-regulated protein kinase, has been shown recently to be part of the CCR4 transcriptional regulatory complex in the yeast Saccharomyces cerevisiae. We report here that temperature-sensitive (ts) dhf2-2 mutant cells can be rescued by overexpression of CDC14, which encodes a dual-specificity protein phosphatase, when grown on glucose-containing medium, as reported previously, but not on galactose. Screening of two S. cerevisiae cDNA libraries led to the identification of CBT1 as a gene which, if overexpressed simultaneously with CDC14, results in the rescue of the dbJ2-2 mutation at restrictive temperature on galactose-based medium, as well as on media containing non-fermentable carbon sources such as glycerol, lactate and acetate. Cbt1 has been shown previously to be essential for formation of cytochrome b in the mitochondria. On the other hand, the ts defects of ccr4delta and caf1delta mutants on glycerol medium at 37 degrees C (Ccr4 and Caf1/Pop2 are two other members of the CCR4 complex) could not be complemented by simultaneous overexpression of CBT1 and CDC14. CCR4 and CAF1 have been shown to play an essential role in activating the expression of genes for non-fermentative growth. Our results strongly suggest that, within the CCR4 complex, Dbf2 is directly involved in some mitochondrial function that depends on cytochrome b or on one of its main regulators, Cbt1. Therefore, Dbf2 may be required not only during mitosis but also during growth on non-fermentable carbon sources.
Dbf2是一种细胞周期调控蛋白激酶,最近研究表明它是酿酒酵母中CCR4转录调控复合体的一部分。我们在此报告,温度敏感型(ts)dhf2-2突变细胞在含葡萄糖的培养基上生长时,如先前报道的那样,可通过过表达编码双特异性蛋白磷酸酶的CDC14来挽救,但在半乳糖培养基上则不能。对两个酿酒酵母cDNA文库的筛选导致鉴定出CBT1基因,若该基因与CDC14同时过表达,则可在基于半乳糖的培养基以及含有甘油、乳酸和乙酸等非发酵碳源的培养基上,在限制温度下挽救dbJ2-2突变。先前已证明Cbt1对于线粒体中细胞色素b 的形成至关重要。另一方面,ccr4delta和caf1delta突变体在37℃甘油培养基上的ts缺陷(Ccr4和Caf1/Pop2是CCR4复合体的另外两个成员)不能通过同时过表达CBT1和CDC14来互补。已证明CCR4和CAF1在激活非发酵生长相关基因的表达中起重要作用。我们的结果强烈表明,在CCR4复合体内,Dbf2直接参与了某些依赖于细胞色素b或其主要调节因子之一Cbt1的线粒体功能。因此,Dbf2可能不仅在有丝分裂期间需要,而且在以非发酵碳源生长期间也需要。