The basal bodies of Chlamydomonas reinhardtii. Formation from probasal bodies, isolation, and partial characterization.
The basal bodies of Chlamydomonas reinhardtii. Formation from probasal bodies, isolation, and partial characterization.
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DOI：
10.1083/jcb.65.1.65
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发表时间：
1975-04
期刊：
影响因子：
--
通讯作者：
Gould RR
中科院分区：
文献类型：
--
作者：
Gould RR
利用无细胞壁菌株cw 15研究了单细胞双鞭毛绿色衣藻Chlamydiumreinhardtii的基体组装和组成。在EDTA的存在下，两个鞭毛轴丝仍然连接到它们的基体，而整个基体轴丝复合物与细胞体分离，没有细胞裂解，用聚乙二醇-400处理。然后在不存在EDTA的情况下从基体中去除轴丝，留下完整的基体对，不受来自细胞其他区域的颗粒污染。分离的细胞器在十二烷基硫酸钠-尿素聚丙烯酰胺凝胶上产生多条带，其中包括两条与鞭毛微管蛋白共沉淀的微管蛋白带。电镜下观察原基底体的形成。将同步细胞裂解，离心到碳包被的网格上，并用铂负染色或阴影。每个细胞的两个原基体在有丝分裂后不久出现薄的“环”，在薄切片中不可见，每个原基体由九个基本的三联微管组成。每个环带保持连接到一个成熟的基体的几个细丝直径约60，并坚持整个间期，直到下一个细胞分裂。然后它延伸成一个成熟的细胞器。结果恢复了基体成核组装的可能性。
The assembly and composition of basal bodies was investigated in the single-celled, biflagellate green alga, Chlamydomonas reinhardtii, using the cell wall-less strain, cw15. In the presence of EDTA, both flagellar axonemes remained attached to their basal bodies while the entire basal body-axoneme complex was separated from the cell body, without cell lysis, by treatment with polyethylene glycol-400. The axonemes were then removed from the basal bodies in the absence of EDTA, leaving intact basal body pairs, free from particulate contamination from other regions of the cell. The isolated organelles produced several bands on sodium dodecyl sulfate-urea polyacrylamide gels, including two tubilin bands which co-electrophoresed with flagellar tubulin. The formation of probasal bodies was observed by electron microscopy of whole mount preparations. Synchronous cells were lysed, centrifuged onto carbon-coated grids, and either negatively stained or shadowed with platinum. The two probasal bodies of each cell appeared shortly after mitosis as thin "annuli," not visible in thin sections, each consisting of nine rudimentary triplet microtubules. Each annulus remained attached to one of the mature basal bodies by several filaments about 60 in diameter, and persisted throughout interphase until just before the next cell division. It then elongated into a mature organelle. The results revive the possibility of the nucleated assembly of basal bodies.