Hoyt M A, He L, Totis L, Saunders W S
Department of Biology, Johns Hopkins University, Baltimore, Maryland 21218.
Genetics. 1993 Sep;135(1):35-44. doi: 10.1093/genetics/135.1.35.
The kinesin-related products of the CIN8 and KIP1 genes of Saccharomyces cerevisiae redundantly perform an essential function in mitosis. The action of either gene-product is required for an outwardly directed force that acts upon the spindle poles. We have selected mutations that suppress the temperature-sensitivity of a cin8-temperature-sensitive kip1-delta strain. The extragenic suppressors analyzed were all found to be alleles of the KAR3 gene. KAR3 encodes a distinct kinesin-related protein whose action antagonizes Cin8p/Kip1p function. All seven alleles analyzed were altered within the region of KAR3 that encodes the putative force-generating (or "motor") domain. These mutations also suppressed the inviability associated with the cin8-delta kip1-delta genotype, a property not shared by a deletion of KAR3. Other properties of the suppressing alleles revealed that they were not null for function. Six of the seven were unaffected for the essential karyogamy and meiosis properties of KAR3 and the seventh was dominant for the suppressing trait. Our findings suggest that despite an antagonistic relationship between Cin8p/Kip1p and Kar3p, aspects of their mitotic roles may be similar.
酿酒酵母CIN8和KIP1基因的驱动蛋白相关产物在有丝分裂中冗余地执行一项基本功能。纺锤体极上向外作用的力需要任一基因产物的作用。我们筛选出了能抑制cin8温度敏感型kip1-δ菌株温度敏感性的突变。分析的基因外抑制子均被发现是KAR3基因的等位基因。KAR3编码一种独特的驱动蛋白相关蛋白,其作用拮抗Cin8p/Kip1p的功能。分析的所有七个等位基因在KAR3编码假定的力产生(或“马达”)结构域的区域内发生了改变。这些突变还抑制了与cin8-δ kip1-δ基因型相关的 inviability,而KAR3缺失则不具有此特性。抑制等位基因的其他特性表明它们并非功能缺失。七个等位基因中的六个对KAR3的基本核融合和减数分裂特性没有影响,第七个在抑制特性方面占主导。我们的研究结果表明,尽管Cin8p/Kip1p和Kar3p之间存在拮抗关系,但它们在有丝分裂中的作用方面可能相似。