Yu G, Li J, Young D
Department of Medical Biochemistry, University of Calgary Health Sciences Centre, Alberta, Canada.
Gene. 1994 Dec 30;151(1-2):215-20. doi: 10.1016/0378-1119(94)90659-9.
We have isolated 16 independent Schizosaccharomyces pombe cDNA clones that suppress the temperature-sensitive (ts) phenotype of a Saccharomyces cerevisiae strain containing the dominant-negative RAS2val19ala22 allele. Fourteen of these cDNAs encode Sz. pombe Ras1. The other two clones encode the C-terminal region of a protein we have named Pka1. We have cloned the pka1 gene from a Sz. pombe genomic library. It contains an uninterrupted open reading frame encoding a 512-amino-acid (aa) protein. The C-terminal region (aa 200-512) of Pka1 is 51-63% identical to cAMP-dependent protein kinase (Pka) catalytic subunits from other eukaryotes. Production of Pka1 suppresses the ts phenotypes exhibited by Sa. cerevisiae ras1-ras2ts or cyr1ts strains. Furthermore, overproduction of Pka1 in Sz. pombe results in a sterile phenotype and an abnormal morphology similar to that exhibited by cells in which the cAMP pathway is constitutively activated. These observations suggest that pka1 encodes the Sz. pombe Pka catalytic subunit.
我们分离出了16个独立的粟酒裂殖酵母cDNA克隆，它们可抑制含有显性负性RAS2val19ala22等位基因的酿酒酵母菌株的温度敏感（ts）表型。其中14个cDNA编码粟酒裂殖酵母Ras1。另外两个克隆编码一种我们命名为Pka1的蛋白质的C末端区域。我们从粟酒裂殖酵母基因组文库中克隆了pka1基因。它包含一个编码512个氨基酸（aa）蛋白质的不间断开放阅读框。Pka1的C末端区域（第200 - 512位氨基酸）与其他真核生物的环磷酸腺苷依赖性蛋白激酶（Pka）催化亚基有51% - 63%的同一性。Pka1的产生抑制了酿酒酵母ras1 - ras2ts或cyr1ts菌株所表现出的ts表型。此外，在粟酒裂殖酵母中过量表达Pka1会导致不育表型和异常形态，类似于环磷酸腺苷途径组成性激活的细胞所表现出的形态。这些观察结果表明pka1编码粟酒裂殖酵母的Pka催化亚基。
