Silencing of a putative inner arm dynein heavy chain results in flagellar immotility in Trypanosoma brucei.
Silencing of a putative inner arm dynein heavy chain results in flagellar immotility in Trypanosoma brucei.
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DOI：
10.1016/j.molbiopara.2010.09.005
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发表时间：
2011-01
影响因子：
1.5
通讯作者：
Klingbeil, Michele M.
中科院分区：
文献类型：
--
作者：
Springer, Amy L.;Bruhn, David F.;Kinzel, Kathryn W.;Rosenthal, Noel F.;Zukas, Randi;Klingbeil, Michele M.
布氏锥虫鞭毛控制运动，对细胞极性和分裂至关重要。锥虫运动的独特功能表明，在这种生物体中的鞭毛节拍调节是不寻常的，值得研究。鞭毛轴丝，运动所需的，具有高度保守的真核生物之间的结构。在轴丝内臂复合体中的几种动力蛋白中，动力蛋白f被认为控制鞭毛的波形形状。霸王预测编码动力蛋白f α重链的布氏杆菌基因TbDNAH 10在原环T中使用RNA干扰沉默。布氏细胞这导致鞭毛不动，除了尖端偶尔轻微抽搐外，没有运动。细胞生长显著减慢，并且发现细胞成大簇。对沉默培养物的显微镜分析显示，许多细胞具有分离的鞭毛，有时缠绕在多个细胞之间。DAPI染色显示错误定位动质体和多核细胞的频率增加，表明这些细胞在细胞周期早期阶段经历破坏，可能继发于运动缺陷。TEM图像显示明显正常的轴丝和内部臂结构没有明显的缺陷。这项研究表明，使用RNAi作为一种有效的方法来研究非常大的基因，如动力蛋白重链（HC），这些动力蛋白敲低的不运动表型表明，一个完整的内臂是必要的鞭毛跳动在T。布鲁塞。由于类似的突变体在衣原体reinhardtii保留运动，这种表型可能反映了两种生物体之间的运动和/或动力蛋白组装的要求的差异，这些比较研究将有助于阐明鞭毛节拍调节的机制。
The Trypanosoma brucei flagellum controls motility and is crucial for cell polarity and division. Unique features of trypanosome motility suggest that flagellar beat regulation in this organism is unusual and worthy of study. The flagellar axoneme, required for motility, has a structure that is highly conserved among eukaryotes. Of the several dyneins in the axonemal inner arm complex, dynein f is thought to control flagellar waveform shape. A T. brucei gene predicted to encode the dynein f alpha heavy chain, TbDNAH10, was silenced using RNA interference in procyclic T. brucei cells. This resulted in immotile flagella, showing no movement except for occasional slight twitches at the tips. Cell growth slowed dramatically and cells were found in large clusters. Microscopic analysis of silenced cultures showed many cells with detached flagella, sometimes entangled between multiple cells. DAPI staining showed an increased frequency of mis-positioned kinetoplasts and multinucleate cells, suggesting that these cells experience disruption at an early cell cycle stage, probably secondary to the motility defect. TEM images showed apparently normal axonemes and no discernable defects in inner arm structure. This study demonstrates use of RNAi as an effective method to study very large genes such as dynein heavy chains (HCs), and the immotility phenotype of these dynein knockdowns suggests that an intact inner arm is necessary for flagellar beating in T. brucei. Since analogous mutants in Chlamydomonas reinhardtii retain motility, this phenotype likely reflects differences in requirements for motility and/or dynein assembly between the two organisms and these comparative studies will help elucidate the mechanisms of flagellar beat regulation.
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DOI：
10.1083/jcb.112.3.441
发表时间：
1991-02
期刊：
The Journal of cell biology
影响因子：
--
作者：
Kamiya R;Kurimoto E;Muto E
通讯作者：
Muto E
一个缺乏外动臂的运动衣原体鞭毛突变体。
影响因子：
7.8
作者：
Mitchell, D R;Rosenbaum, J L
通讯作者：
Rosenbaum, J L
影响因子：
11.4
作者：
Kohl, L;Robinson, D;Bastin, P
通讯作者：
Bastin, P
影响因子：
--
作者：
BROKAW, CJ;LUCK, DJL
通讯作者：
LUCK, DJL
影响因子：
--
作者：
Chandler, Julian;Vandoros, Anthula V.;Klingbeil, Michele M.
通讯作者：
Klingbeil, Michele M.