Roof D M, Meluh P B, Rose M D
Department of Molecular Biology, Princeton University, New Jersey 08544.
J Cell Biol. 1992 Jul;118(1):95-108. doi: 10.1083/jcb.118.1.95.
We identified two new Saccharomyces cerevisiae kinesin-related genes, KIP1 and KIP2, using polymerase chain reaction primers corresponding to highly conserved regions of the kinesin motor domain. Both KIP proteins are expressed in vivo, but deletion mutations conferred no phenotype. Moreover, kip1 kip2 double mutants and a triple mutant with kinesin-related kar3 had no synthetic phenotype. Using a genetic screen for mutations that make KIP1 essential, we identified another gene, KSL2, which proved to be another kinesin-related gene, CIN8. KIP1 and CIN8 are functionally redundant: double mutants arrested in mitosis whereas the single mutants did not. The microtubule organizing centers of arrested cells were duplicated but unseparated, indicating that KIP1 or CIN8 is required for mitotic spindle assembly. Consistent with this role, KIP1 protein was found to colocalize with the mitotic spindle.
我们使用对应于驱动蛋白运动结构域高度保守区域的聚合酶链反应引物，鉴定出两个新的酿酒酵母驱动蛋白相关基因KIP1和KIP2。两种KIP蛋白均在体内表达，但缺失突变未产生表型。此外，kip1 kip2双突变体以及与驱动蛋白相关的kar3三突变体均无合成表型。通过对使KIP1成为必需基因的突变进行遗传筛选，我们鉴定出另一个基因KSL2，结果证明它是另一个驱动蛋白相关基因CIN8。KIP1和CIN8在功能上是冗余的：双突变体在有丝分裂中停滞，而单突变体则不会。停滞细胞的微管组织中心发生了复制但未分离，这表明有丝分裂纺锤体组装需要KIP1或CIN8。与此作用一致，发现KIP1蛋白与有丝分裂纺锤体共定位。
