de Barros Lopes M, Ho J Y, Reed S I
Department of Molecular Biology, Research Institute of Scripps Clinic, La Jolla, California 92037.
Mol Cell Biol. 1990 Jun;10(6):2966-72. doi: 10.1128/mcb.10.6.2966-2972.1990.
Conditional mutations in the genes CDC36 and CDC39 cause arrest in the G1 phase of the Saccharomyces cerevisiae cell cycle at the restrictive temperature. We present evidence that this arrest is a consequence of a mutational activation of the mating pheromone response. cdc36 and cdc39 mutants expressed pheromone-inducible genes in the absence of pheromone and conjugated in the absence of a mating pheromone receptor. On the other hand, cells lacking the G beta subunit or overproducing the G alpha subunit of the transducing G protein that couples the receptor to the pheromone response pathway prevented constitutive activation of the pathway in cdc36 and cdc39 mutants. These epistasis relationships imply that the CDC36 and CDC39 gene products act at the level of the transducing G protein. The CDC36 and CDC39 gene products have a role in cellular processes other than the mating pheromone response. A mating-type heterozygous diploid cell, homozygous for either the cdc36 or cdc39 mutation, does not exhibit the G1 arrest phenotype but arrests asynchronously with respect to the cell cycle. A similar asynchronous arrest was observed in cdc36 and cdc39 cells where the pheromone response pathway had been inactivated by mutations in the transducing G protein. Furthermore, cdc36 and cdc39 mutants, when grown on carbon catabolite-derepressing medium, did not arrest in G1 and did not induce pheromone-specific genes at the restrictive temperature.
CDC36和CDC39基因中的条件突变会导致酿酒酵母细胞周期在限制温度下停滞于G1期。我们提供的证据表明,这种停滞是交配信息素反应的突变激活的结果。cdc36和cdc39突变体在没有信息素的情况下表达信息素诱导型基因,并且在没有交配信息素受体的情况下进行接合。另一方面,缺乏转导G蛋白的Gβ亚基或过量产生其Gα亚基的细胞,该转导G蛋白将受体与信息素反应途径偶联,可阻止cdc36和cdc39突变体中该途径的组成型激活。这些上位性关系表明,CDC36和CDC39基因产物在转导G蛋白水平上起作用。CDC36和CDC39基因产物在除交配信息素反应之外的细胞过程中也发挥作用。对于cdc36或cdc39突变纯合的交配型杂合二倍体细胞,不会表现出G1停滞表型,而是相对于细胞周期异步停滞。在cdc36和cdc39细胞中观察到类似的异步停滞,其中信息素反应途径已因转导G蛋白的突变而失活。此外,cdc36和cdc39突变体在碳源分解代谢去阻遏培养基上生长时,在限制温度下不会停滞于G1期,也不会诱导信息素特异性基因。