Pereira A J, Dalby B, Stewart R J, Doxsey S J, Goldstein L S
Program in Molecular Medicine, University of Massachusetts Medical Center, Worcester 01605, USA.
J Cell Biol. 1997 Mar 10;136(5):1081-90. doi: 10.1083/jcb.136.5.1081.
The kinesin superfamily is a large group of proteins (kinesin-like proteins [KLPs]) that share sequence similarity with the microtubule (MT) motor kinesin. Several members of this superfamily have been implicated in various stages of mitosis and meiosis. Here we report our studies on KLP67A of Drosophila. DNA sequence analysis of KLP67A predicts an MT motor protein with an amino-terminal motor domain. To prove this directly, KLP67A expressed in Escherichia coli was shown in an in vitro motility assay to move MTs in the plus direction. We also report expression analyses at both the mRNA and protein level, which implicate KLP67A in the localization of mitochondria in undifferentiated cell types. In situ hybridization studies of the KLP67A mRNA during embryogenesis and larval central nervous system development indicate a proliferation-specific expression pattern. Furthermore, when affinity-purified anti-KLP67A antisera are used to stain blastoderm embryos, mitochondria in the region of the spindle asters are labeled. These data suggest that KLP67A is a mitotic motor of Drosophila that may have the unique role of positioning mitochondria near the spindle.
驱动蛋白超家族是一大类蛋白质(类驱动蛋白[KLPs]),它们与微管(MT)驱动蛋白驱动蛋白在序列上具有相似性。该超家族的几个成员与有丝分裂和减数分裂的各个阶段有关。在此,我们报告我们对果蝇KLP67A的研究。对KLP67A的DNA序列分析预测其为一种具有氨基末端驱动结构域的MT驱动蛋白。为了直接证明这一点,在体外运动分析中,显示在大肠杆菌中表达的KLP67A能使MT向正方向移动。我们还报告了在mRNA和蛋白质水平的表达分析,这表明KLP67A与未分化细胞类型中线粒体的定位有关。在胚胎发生和幼虫中枢神经系统发育过程中对KLP67A mRNA进行的原位杂交研究表明其具有增殖特异性表达模式。此外,当使用亲和纯化的抗KLP67A抗血清对囊胚期胚胎进行染色时,纺锤体星状体区域的线粒体被标记。这些数据表明,KLP67A是果蝇的一种有丝分裂驱动蛋白,可能具有将线粒体定位在纺锤体附近的独特作用。