Ogawa K, Kamiya R, Wilkerson C G, Witman G B
Department of Cell Biology, National Institute for Basic Biology, Okazaki, Japan.
Mol Biol Cell. 1995 Jun;6(6):685-96. doi: 10.1091/mbc.6.6.685.
Immunological analysis showed that antibodies against the intermediate chains (ICs) IC2 and IC3 of sea urchin outer arm dynein specifically cross-reacted with intermediate chains IC78 and IC69, respectively, of Chlamydomonas outer arm dynein. In contrast, no specific cross-reactivity with any Chlamydomonas outer arm polypeptide was observed using antibody against IC1 of sea urchin outer arm dynein. To learn more about the relationships between the different ICs, overlapping cDNAs encoding all of IC2 and IC3 of sea urchin were isolated and sequenced. Comparison of these sequences with those previously obtained for the Chlamydomonas ICs revealed that, although all four chains are homologous, sea urchin IC2 is much more closely related to Chlamydomonas IC78 (45.8% identity), and sea urchin IC3 is much more closely related to Chlamydomonas IC69 (48.5% identity), than either sea urchin chain is related to the other (23.5% identity). For homologous pairs, the similarities extend throughout the full lengths of the chains. Regions of similarity between all four ICs and the IC (IC74) of cytoplasmic dynein, located in the C-terminal halves of the chains, are due primarily to conservation of the WD repeats present in all of these ICs. This is the first demonstration that structural differences between individual ICs within an outer arm dynein have been highly conserved in the dyneins of distantly related species. The results provide a basis for the subclassification of these chains.
免疫分析表明,针对海胆外臂动力蛋白中间链(ICs)IC2和IC3的抗体分别与衣藻外臂动力蛋白的中间链IC78和IC69发生特异性交叉反应。相比之下,使用针对海胆外臂动力蛋白IC1的抗体未观察到与任何衣藻外臂多肽的特异性交叉反应。为了更多地了解不同中间链之间的关系,分离并测序了编码海胆所有IC2和IC3的重叠cDNA。将这些序列与先前获得的衣藻中间链序列进行比较,结果显示,尽管所有四条链都是同源的,但海胆IC2与衣藻IC78的关系更为密切(同一性为45.8%),海胆IC3与衣藻IC69的关系更为密切(同一性为48.5%),而海胆的两条链之间的关系则较弱(同一性为23.5%)。对于同源对,相似性贯穿链的全长。所有四条中间链与位于链C端一半的胞质动力蛋白的中间链(IC74)之间的相似区域,主要是由于所有这些中间链中存在的WD重复序列的保守性。这是首次证明外臂动力蛋白中单个中间链之间的结构差异在远缘物种的动力蛋白中高度保守。这些结果为这些链的亚分类提供了基础。