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首页> 外文期刊>Cell motility and the cytoskeleton >Transport and arrangement of the outer-dynein-arm docking complex in the flagella of Chlamydomonas mutants that lack outer dynein arms.
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Transport and arrangement of the outer-dynein-arm docking complex in the flagella of Chlamydomonas mutants that lack outer dynein arms.

机译：缺乏外部动力蛋白臂的衣藻突变体鞭毛中的外部动力蛋白臂对接复合体的运输和排列。

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The outer dynein arms of Chlamydomonas flagella are attached to a precise site on the outer doublet microtubules and repeat at a regular interval of 24 nm. This binding is mediated by the outer dynein arm docking complex (ODA-DC), which is composed of three protein subunits. In this study, antibodies against the 83- and 62-kD subunits (DC83 and DC62) of the ODA-DC were used to analyze its state of association with outer arm components within the cytoplasm, and its localization in the axonemes of oda mutants. Immunoprecipitation indicates that DC83 and DC62 are preassembled within the cytoplasm, but that they are not associated with outer arm dynein. Both proteins are lost or greatly diminished in oda1 and oda3, mutants in the structural genes of DC62 and DC83, respectively, demonstrating that their association is necessary for their stable presence in the cytoplasm. Immunoelectron microscopy indicates that DC83 repeats at 24-nm intervals along the length of the doublet microtubules of oda6, which lacks outer arms; thus, outer arm periodicity may be determined by the ODA-DC. Flagellar regeneration and temporary dikaryon experiments indicate that the ODA-DC can be rapidly transported into the flagellum and assembled on the doublet microtubules independently of the outer arms and independently of flagellar growth. Unexpectedly, the intensity of ODA-DC labeling decreased toward the distal ends of axonemes of oda6 but not wild-type cells, suggesting that the outer arms reciprocally contribute to the assembly/stability of the ODA-DC. Copyright 2001 Wiley-Liss, Inc.
机译：衣藻鞭毛的外部动力蛋白臂附着在外部双峰微管上的精确位置，并以24 nm的规则间隔重复。这种结合是由外部动力蛋白对接复合物（ODA-DC）介导的，该复合物由三个蛋白质亚基组成。在这项研究中，使用了针对ODA-DC的83-kD和62-kD亚基（DC83和DC62）的抗体来分析其与细胞质中外臂成分的缔合状态，以及其在oda突变体的轴突中的定位。免疫沉淀表明DC83和DC62在细胞质中预先组装，但与外臂动力蛋白无关。这两种蛋白质分别在dc1和dc83结构基因的oda1和oda3突变体中丢失或大大减少，表明它们的缔合对于它们在细胞质中的稳定存在是必要的。免疫电子显微镜检查表明，DC83沿oda6的双峰微管的长度方向以24 nm的间隔重复，而oda6的微管没有外部臂。因此，外臂周期性可由ODA-DC确定。鞭毛再生和临时双核生物实验表明，ODA-DC可以快速运输到鞭毛中，并独立于外臂和鞭毛生长而组装在双态微管上。出乎意料的是，ODA-DC标记的强度朝着oda6的轴突的远端降低，但没有向野生型细胞下降，这表明外臂对ODA-DC的组装/稳定性起反作用。版权所有2001 Wiley-Liss，Inc.

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