Baum P, Furlong C, Byers B
Proc Natl Acad Sci U S A. 1986 Aug;83(15):5512-6. doi: 10.1073/pnas.83.15.5512.
Saccharomyces cerevisiae strains bearing temperature-sensitive alleles of the cell division cycle gene CDC31 are specifically defective in duplication of the spindle pole body, the microtubule-organizing center of yeast. To define the function encoded by CDC31 more fully, we have isolated genomic clones of the gene by selection for complementation of a temperature-sensitive allele. The locus from which the clone was derived was marked by integration of a nutritional marker and found by meiotic mapping to cosegregate with CDC31. The polypeptide sequence of the open reading frame in the CDC31 gene was determined and compared with the sequences of other known proteins. This revealed significant homology with the calmodulins and other members of the Ca2+-binding protein family. On the basis of comparison with these related proteins, it is evident that the CDC31 gene product has at least two binding sites for Ca2+ and is also homologous with other regions of the calmodulin sequence. We propose that Ca2+ fluxes within the yeast cell play a key role in spindle pole body duplication and consequently in the organization of the microtubule arrays.
携带细胞分裂周期基因CDC31温度敏感等位基因的酿酒酵母菌株在纺锤体极体(酵母的微管组织中心)的复制过程中存在特异性缺陷。为了更全面地定义CDC31编码的功能,我们通过选择对温度敏感等位基因进行互补来分离该基因的基因组克隆。克隆所源自的位点通过营养标记的整合进行标记,并通过减数分裂定位发现与CDC31共分离。确定了CDC31基因中开放阅读框的多肽序列,并与其他已知蛋白质的序列进行了比较。这揭示了与钙调蛋白和Ca2+结合蛋白家族其他成员的显著同源性。基于与这些相关蛋白质的比较,很明显CDC31基因产物至少有两个Ca2+结合位点,并且在钙调蛋白序列的其他区域也具有同源性。我们提出,酵母细胞内的Ca2+通量在纺锤体极体复制以及因此在微管阵列的组织中起关键作用。