Rischitor Patricia E, Konzack Sven, Fischer Reinhard
Department of Microbiology, University of Marburg, D-35043 Marburg, Germany.
Eukaryot Cell. 2004 Jun;3(3):632-45. doi: 10.1128/EC.3.3.632-645.2004.
Kinesins are motor proteins which are classified into 11 different families. We identified 11 kinesin-like proteins in the genome of the filamentous fungus Aspergillus nidulans. Relatedness analyses based on the motor domains grouped them into nine families. In this paper, we characterize KipB as a member of the Kip3 family of microtubule depolymerases. The closest homologues of KipB are Saccharomyces cerevisiae Kip3 and Schizosaccharomyces pombe Klp5 and Klp6, but sequence similarities outside the motor domain are very low. A disruption of kipB demonstrated that it is not essential for vegetative growth. kipB mutant strains were resistant to high concentrations of the microtubule-destabilizing drug benomyl, suggesting that KipB destabilizes microtubules. kipB mutations caused a failure of spindle positioning in the cell, a delay in mitotic progression, an increased number of bent mitotic spindles, and a decrease in the depolymerization of cytoplasmic microtubules during interphase and mitosis. Meiosis and ascospore formation were not affected. Disruption of the kipB gene was synthetically lethal in combination with the temperature-sensitive mitotic kinesin motor mutation bimC4, suggesting an important but redundant role of KipB in mitosis. KipB localized to cytoplasmic, astral, and mitotic microtubules in a discontinuous pattern, and spots of green fluorescent protein moved along microtubules toward the plus ends.
驱动蛋白是一类运动蛋白,可分为11个不同的家族。我们在丝状真菌构巢曲霉的基因组中鉴定出了11种驱动蛋白样蛋白。基于运动结构域的相关性分析将它们分为9个家族。在本文中,我们将KipB鉴定为微管解聚酶Kip3家族的一员。KipB最接近的同源物是酿酒酵母的Kip3以及粟酒裂殖酵母的Klp5和Klp6,但在运动结构域之外的序列相似性非常低。kipB的缺失表明它对于营养生长并非必需。kipB突变菌株对高浓度的微管不稳定药物苯菌灵具有抗性,这表明KipB会使微管不稳定。kipB突变导致细胞中纺锤体定位失败、有丝分裂进程延迟、弯曲的有丝分裂纺锤体数量增加,以及在间期和有丝分裂期间细胞质微管解聚减少。减数分裂和子囊孢子形成不受影响。kipB基因的缺失与温度敏感的有丝分裂驱动蛋白运动突变bimC4组合时具有合成致死性,这表明KipB在有丝分裂中具有重要但冗余的作用。KipB以不连续的模式定位于细胞质、星体和有丝分裂微管,绿色荧光蛋白斑点沿着微管向正端移动。