Regulation of flagellar dynein by phosphorylation of a 138-kD inner arm dynein intermediate chain.
Regulation of flagellar dynein by phosphorylation of a 138-kD inner arm dynein intermediate chain.
复制标题
DOI：
10.1083/jcb.136.1.167
复制
发表时间：
1997-01-13
期刊：
影响因子：
--
通讯作者：
Sale WS
中科院分区：
文献类型：
--
作者：
Habermacher G;Sale WS
理解纤毛和鞭毛运动的挑战之一是确定局部调节动力蛋白驱动的微管滑动的机制。我们最近的研究表明，鞭毛衣单胞菌的微管滑动受磷酸化调节。然而，调控蛋白仍然未知。在这里，我们确定了138-kD内臂动力蛋白I1的中间链是调节运动所需的关键磷酸化蛋白。这个结论是建立在三种不同实验方法的结果之上的。首先，遗传分析和功能分析表明，通过磷酸化调控微管滑动需要内臂动力蛋白11。其次，体外磷酸化表明I1的138-kD中间链是唯一被磷酸化的亚基。第三，体外重构表明138-kD中间链的磷酸化和去磷酸化分别抑制和恢复野生型微管滑动。我们得出结论，I1的138-kD中间链磷酸化的变化调节动力蛋白驱动的微管滑动。此外，基于这些和其他数据，我们预测I1活性的调节参与了鞭毛波形的调制。
One of the challenges in understanding ciliary and flagellar motility is determining the mechanisms that locally regulate dynein-driven microtubule sliding. Our recent studies demonstrated that microtubule sliding, in Chlamydomonas flagella, is regulated by phosphorylation. However, the regulatory proteins remain unknown. Here we identify the 138-kD intermediate chain of inner arm dynein I1 as the critical phosphoprotein required for regulation of motility. This conclusion is founded on the results of three different experimental approaches. First, genetic analysis and functional assays revealed that regulation of microtubule sliding, by phosphorylation, requires inner arm dynein I1. Second, in vitro phosphorylation indicated the 138-kD intermediate chain of I1 is the only phosphorylated subunit. Third, in vitro reconstitution demonstrated that phosphorylation and dephosphorylation of the 138-kD intermediate chain inhibits and restores wild-type microtubule sliding, respectively. We conclude that change in phosphorylation of the 138-kD intermediate chain of I1 regulates dynein-driven microtubule sliding. Moreover, based on these and other data, we predict that regulation of I1 activity is involved in modulation of flagellar waveform.