Bridge A J, Morphew M, Bartlett R, Hagan I M
School of Biological Sciences, University of Manchester, Manchester, M13 9PT UK.
Genes Dev. 1998 Apr 1;12(7):927-42. doi: 10.1101/gad.12.7.927.
During fission yeast mitosis, the duplicated spindle pole bodies (SPBs) nucleate microtubule arrays that interdigitate to form the mitotic spindle. cut12.1 mutants form a monopolar mitotic spindle, chromosome segregation fails, and the mutant undergoes a lethal cytokinesis. The cut12(+) gene encodes a novel 62-kD protein with two predicted coiled coil regions, and one consensus phosphorylation site for p34(cdc2) and two for MAP kinase. Cut12 is localized to the SPB throughout the cell cycle, predominantly around the inner face of the interphase SPB, adjacent to the nucleus. cut12(+) is allelic to stf1(+); stf1.1 is a gain-of-function mutation bypassing the requirement for the Cdc25 tyrosine phosphatase, which normally dephosphorylates and activates the p34(cdc2)/cyclin B kinase to promote the onset of mitosis. Expressing a cut12(+) cDNA carrying the stf1.1 mutation also suppressed cdc25.22. The spindle defect in cut12.1 is exacerbated by the cdc25.22 mutation, and stf1.1 cells formed defective spindles in a cdc25.22 background at high temperatures. We propose that Cut12 may be a regulator or substrate of the p34(cdc2) mitotic kinase.
在裂殖酵母有丝分裂过程中,复制后的纺锤体极体(SPB)形成相互交错的微管阵列以构成有丝分裂纺锤体。cut12.1突变体形成单极有丝分裂纺锤体,染色体分离失败,且该突变体经历致死性胞质分裂。cut12(+)基因编码一种新型的62-kD蛋白,具有两个预测的卷曲螺旋区域,一个p34(cdc2)的共有磷酸化位点和两个丝裂原活化蛋白激酶的共有磷酸化位点。Cut12在整个细胞周期中定位于SPB,主要围绕间期SPB的内表面,与细胞核相邻。cut12(+)与stf1(+)等位;stf1.1是一个功能获得性突变,绕过了对Cdc25酪氨酸磷酸酶的需求,Cdc25酪氨酸磷酸酶通常使p34(cdc2)/细胞周期蛋白B激酶去磷酸化并激活它,以促进有丝分裂的开始。表达携带stf1.1突变的cut12(+) cDNA也能抑制cdc25.22。cdc25.22突变会加剧cut12.1中的纺锤体缺陷,并且stf1.1细胞在高温下的cdc25.22背景中形成有缺陷的纺锤体。我们提出Cut12可能是p34(cdc2)有丝分裂激酶的调节因子或底物。