Toda T, Shimanuki M, Yanagida M
Department of Biophysics, Faculty of Science, Kyoto University, Japan.
EMBO J. 1993 May;12(5):1987-95. doi: 10.1002/j.1460-2075.1993.tb05848.x.
Two novel protein kinase C (PKC)-like genes, pck1+ and pck2+ were isolated from fission yeast by PCR. Both contain common domains of PKC-related molecules, but lack a putative Ca(2+)-binding domain so that they may belong to the nPKC group. Gene disruption of pck1+ and pck2+ establishes that they share an overlapping essential function for cell viability. Cells of a single pck2 deletion display severe defects in cell shape; they are irregular and sometimes pear-like instead of cylindrical. In contrast, the induced overexpression of pck2+ is lethal, producing multiseptated and branched cells. These results suggest that fission yeast PKC-like genes are involved in the polarity of cell growth control. We show that pck2 is allelic to sts6, a locus we have previously identified by its supersensitivity to staurosporine, a potent protein kinase inhibitor [Toda et al. (1991) Genes Dev., 5, 60-73]. In addition, the lethal overexpression of pck2+ can be suppressed by staurosporine, indicating that fission yeast pck1 and pck2 are molecular targets of this inhibitor.
通过聚合酶链反应（PCR）从裂殖酵母中分离出两个新的蛋白激酶C（PKC）样基因，即pck1 +和pck2 +。两者都含有PKC相关分子的共同结构域，但缺乏假定的Ca(2 +)结合结构域，因此它们可能属于非典型PKC（nPKC）组。对pck1 +和pck2 +进行基因敲除表明，它们对于细胞活力具有重叠的基本功能。单个pck2缺失的细胞在细胞形态上表现出严重缺陷；它们不规则，有时呈梨形而非圆柱形。相反，pck2 +的诱导过表达是致死性的，会产生多隔和分支的细胞。这些结果表明，裂殖酵母PKC样基因参与细胞生长控制的极性。我们发现pck2与sts6等位，sts6是我们先前通过其对强效蛋白激酶抑制剂星形孢菌素的超敏感性而鉴定出的一个基因座[Toda等人（1991年），《基因与发育》，5，60 - 73]。此外，pck2 +的致死性过表达可被星形孢菌素抑制，这表明裂殖酵母中的pck1和pck2是该抑制剂的分子靶点。
