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首页> 外文期刊>Cell motility and the cytoskeleton >High speed sliding of axonemal microtubules produced by outer arm dynein.
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High speed sliding of axonemal microtubules produced by outer arm dynein.

机译：外臂动力蛋白产生的轴突微管高速滑动。

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摘要

To study dynein arm activity at high temporal resolution, axonemal sliding was measured field by field for wild type and dynein arm mutants of Tetrahymena thermophila. For wt SB255 cells, when the rate of data acquisition was 60 fps, about 5x greater than previously published observations, sliding was observed to be discontinuous with very high velocity sliding (average 196 microm/sec) for a few msec (1 or 2 fields) followed by a pause of several fields. The sliding velocities measured were an order of magnitude greater than rates previously measured by video analysis. However, when the data were analyzed at 12 fps for the same axonemes, consistent with previous observations, sliding was linear as the axonemes extended several times their original length with an average velocity of approximately 10 microm/sec. The pauses or stops occurred at approximately 200 and 300% of the initial length, suggesting that dynein arms on one axonemal doublet were initially active to the limit of extension, and then thearms on the next doublet became activated. In contrast, in a mutant where OADs are missing, sliding observed at 60 fps was continuous and slow (5 microm/sec), as opposed to the discontinuous high-velocity sliding of SB255 and of the mutant at the permissive temperature where OADs are present. High-velocity step-wise sliding was also present in axonemes from an inner arm dynein mutant (KO6). These results indicate that the high-speed discontinuous pattern of sliding is produced by the mechanochemical activity of outer arm dynein. The rate of sliding is consistent with a low duty ratio of the outer arm dynein and with the operation of each arm along a doublet once per beat.
机译：为了研究高时间分辨率下的达因臂活性，对嗜热四膜虫的野生型和达因臂突变体逐场测量了轴突滑动。对于wt SB255电池，当数据采集速率为60 fps，比以前发表的观察结果大5倍时，观察到滑动是不连续的，并且具有很高的速度滑动（平均196 microm / sec），持续了几毫秒（1或2个场） ），然后暂停几个字段。测得的滑动速度比以前通过视频分析测得的速率大一个数量级。但是，当以12 fps分析相同轴突的数据时，与以前的观察结果一致，滑移是线性的，因为轴突以其原始长度扩展了几倍，平均速度约为10微米/秒。停顿或停止发生在初始长度的大约200％和300％处，这表明一个轴突双联体上的动力蛋白臂最初处于活动状态，直到伸展的极限，然后下一个双联体上的动力臂被激活。相反，在缺少OAD的突变体中，以60 fps观察到的滑动是连续且缓慢的（5微米/秒），这与SB255和突变体在存在OAD的容许温度下不连续的高速滑动相反。内臂动力蛋白突变体（KO6）的轴突中也存在高速逐步滑动。这些结果表明，高速不连续滑动是由外臂动力蛋白的机械化学活性产生的。滑动速度与外臂动力蛋白的低占空比以及每个拍子沿双音调的每个臂的操作一致。

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