Miller J M, Wang W, Balczon R, Dentler W L
Department of Physiology and Cell Biology, University of Kansas, Lawrence 66045-2106.
J Cell Biol. 1990 Mar;110(3):703-14. doi: 10.1083/jcb.110.3.703.
Structures that cap the plus ends of microtubules may be involved in the regulation of their assembly and disassembly. Growing and disassembling microtubules in the mitotic apparatus are capped by kinetochores and ciliary and flagellar microtubules are capped by the central microtubule cap and distal filaments. To compare the ciliary caps with kinetochores, isolated Tetrahymena cilia were stained with CREST (Calcinosis/phenomenon esophageal dysmotility, sclerodactyly, telangiectasia) antisera known to stain kinetochores. Immunofluorescence microscopy revealed that a CREST antiserum stained the distal tips of cilia that contained capping structures but did not stain axonemes that lacked capping structures. Both Coomassie blue-stained gels and Western blots probed with CREST antiserum revealed that a 97-kD antigen copurifies with the capping structures. Affinity-purified antibodies to the 97-kD ciliary protein stained the tips of cap-containing Tetrahymena cilia and the kinetochores in HeLa, Chinese hamster ovary, and Indian muntjak cells. These results suggest that at least one polypeptide found in the kinetochore is present in ciliary microtubule capping structures and that there may be a structural and/or functional homology between these structures that cap the plus ends of microtubules.
覆盖微管正端的结构可能参与其组装和解聚的调节。有丝分裂装置中正在生长和解聚的微管由动粒覆盖,而纤毛和鞭毛微管则由中央微管帽和远端丝覆盖。为了将纤毛帽与动粒进行比较,用已知可对动粒进行染色的抗着丝点血清(钙质沉着/食管运动障碍、指(趾)硬皮病、毛细血管扩张现象)对分离的四膜虫纤毛进行染色。免疫荧光显微镜检查显示,抗着丝点血清可对含有覆盖结构的纤毛远端进行染色,但不能对缺乏覆盖结构的轴丝进行染色。考马斯亮蓝染色凝胶和用抗着丝点血清进行的蛋白质印迹分析均显示,一种97-kD抗原与覆盖结构共同纯化。针对97-kD纤毛蛋白的亲和纯化抗体可对含有帽的四膜虫纤毛尖端以及HeLa细胞、中国仓鼠卵巢细胞和印度麂细胞中的动粒进行染色。这些结果表明,在着丝粒中发现的至少一种多肽存在于纤毛微管覆盖结构中,并且这些覆盖微管正端的结构之间可能存在结构和/或功能同源性。