Kawamukai M, Ferguson K, Wigler M, Young D
Cold Spring Harbor Laboratory, New York 11724.
Cell Regul. 1991 Feb;2(2):155-64. doi: 10.1091/mbc.2.2.155.
The adenylyl cyclase gene, cyr1, of Schizosaccharomyces pombe has been cloned. We have begun an analysis of the function and regulation of adenylyl cyclase by disrupting this gene and by over-expressing all or parts of this gene in various strains. cyr1- strains are viable and contain no measurable cyclic AMP. They conjugate and sporulate under conditions that normally inhibit wild-type strains. Strains containing the cyr1 coding sequences transcribed from the strong adh1 promoter contain greatly elevated adenylyl cyclase activity, as measured in vitro, but only modestly elevated cAMP levels. Such strains conjugate and sporulate less frequently than wild-type cells upon nutrient limitation. Strains which carry the wild-type cyr1 gene but that also express high levels of the amino terminal domain of adenylyl cyclase behave much like cyr1-strains, suggesting that the amino terminal domain can bind a positive regulator. A protein that copurifies with the adenylyl cyclase of S. pombe cross-reacts to antiserum raised against the S. cerevisiae adenylyl cyclase-associated regulatory protein, CAP.
粟酒裂殖酵母的腺苷酸环化酶基因cyr1已被克隆。我们通过破坏该基因以及在不同菌株中过表达该基因的全部或部分来开始对腺苷酸环化酶的功能和调控进行分析。cyr1基因缺失菌株可以存活，且不含可检测到的环磷酸腺苷（cAMP）。它们在通常会抑制野生型菌株的条件下进行接合和孢子形成。含有从强adh1启动子转录的cyr1编码序列的菌株，经体外测定，其腺苷酸环化酶活性大幅升高，但cAMP水平仅适度升高。在营养限制条件下，这类菌株的接合和孢子形成频率低于野生型细胞。携带野生型cyr1基因但也高水平表达腺苷酸环化酶氨基末端结构域的菌株，其行为与cyr1基因缺失菌株非常相似，这表明氨基末端结构域可以结合一种正向调节因子。一种与粟酒裂殖酵母腺苷酸环化酶共纯化的蛋白质，能与针对酿酒酵母腺苷酸环化酶相关调节蛋白CAP产生的抗血清发生交叉反应。
