Salisbury J L, Baron A T, Sanders M A
Center for NeuroSciences, School of Medicine, Case Western Reserve University, Cleveland, Ohio 44106.
J Cell Biol. 1988 Aug;107(2):635-41. doi: 10.1083/jcb.107.2.635.
Monoclonal and polyclonal antibodies raised against algal centrin, a protein of algal striated flagellar roots, were used to characterize the occurrence and distribution of this protein in interphase and mitotic Chlamydomonas cells. Chlamydomonas centrin, as identified by Western immunoblot procedures, is a low molecular (20,000-Mr) acidic protein. Immunofluorescence and immunogold labeling demonstrates that centrin is a component of the distal fiber. In addition, centrin-based flagellar roots link the flagellar apparatus to the nucleus. Two major descending fibers extend from the basal bodies toward the nucleus; each descending fiber branches several times giving rise to 8-16 fimbria which surround and embrace the nucleus. Immunogold labeling indicates that these fimbria are juxtaposed to the outer nuclear envelope. Earlier studies have demonstrated that the centrin-based linkage between the flagellar apparatus and the nucleus is contractile, both in vitro and in living Chlamydomonas cells (Wright, R. L., J. Salisbury, and J. Jarvik. 1985. J. Cell Biol. 101:1903-1912; Salisbury, J. L., M. A. Sanders, and L. Harpst. 1987. J. Cell Biol. 105:1799-1805). Immunofluorescence studies show dramatic changes in distribution of the centrin-based system during mitosis that include a transient contraction at preprophase; division, separation, and re-extension during prophase; and a second transient contraction at the metaphase/anaphase boundary. These observations suggest a fundamental role for centrin in motile events during mitosis.
针对藻类中心蛋白(一种藻类横纹鞭毛根部的蛋白质)制备的单克隆抗体和多克隆抗体,被用于表征该蛋白在衣藻细胞间期和有丝分裂期的出现情况和分布。通过蛋白质免疫印迹法鉴定,衣藻中心蛋白是一种低分子量(20,000道尔顿)的酸性蛋白。免疫荧光和免疫金标记表明,中心蛋白是远端纤维的一个组成部分。此外,基于中心蛋白的鞭毛根部将鞭毛装置与细胞核相连。两条主要的下行纤维从基体向细胞核延伸;每条下行纤维多次分支,产生8 - 16条纤毛,这些纤毛围绕并包裹着细胞核。免疫金标记表明这些纤毛与核外膜并列。早期研究表明,无论是在体外还是在活的衣藻细胞中,鞭毛装置与细胞核之间基于中心蛋白的连接都是可收缩的(赖特,R.L.,J.索尔兹伯里,和J.贾维克。1985年。《细胞生物学杂志》101:1903 - 1912;索尔兹伯里,J.L.,M.A.桑德斯,和L.哈普斯特。1987年。《细胞生物学杂志》105:1799 - 1805)。免疫荧光研究显示,在有丝分裂期间,基于中心蛋白的系统分布发生了显著变化,包括前期前的短暂收缩;前期的分裂、分离和重新延伸;以及中期/后期边界处的第二次短暂收缩。这些观察结果表明中心蛋白在有丝分裂期间的运动事件中起着重要作用。