Cell Architecture Laboratory, Center for Frontier Research, National Institute of Genetics, Mishima 411-8540, Japan.
Proc Natl Acad Sci U S A. 2011 Jan 4;108(1):137-42. doi: 10.1073/pnas.1013275108. Epub 2010 Dec 20.
The centrosome is generally maintained at the center of the cell. In animal cells, centrosome centration is powered by the pulling force of microtubules, which is dependent on cytoplasmic dynein. However, it is unclear how dynein brings the centrosome to the cell center, i.e., which structure inside the cell functions as a substrate to anchor dynein. Here, we provide evidence that a population of dynein, which is located on intracellular organelles and is responsible for organelle transport toward the centrosome, generates the force required for centrosome centration in Caenorhabditis elegans embryos. By using the database of full-genome RNAi in C. elegans, we identified dyrb-1, a dynein light chain subunit, as a potential subunit involved in dynein anchoring for centrosome centration. DYRB-1 is required for organelle movement toward the minus end of the microtubules. The temporal correlation between centrosome centration and the net movement of organelle transport was found to be significant. Centrosome centration was impaired when Rab7 and RILP, which mediate the association between organelles and dynein in mammalian cells, were knocked down. These results indicate that minus end-directed transport of intracellular organelles along the microtubules is required for centrosome centration in C. elegans embryos. On the basis of this finding, we propose a model in which the reaction forces of organelle transport generated along microtubules act as a driving force that pulls the centrosomes toward the cell center. This is the first model, to our knowledge, providing a mechanical basis for cytoplasmic pulling force for centrosome centration.
中心体通常位于细胞的中心。在动物细胞中,中心体的集中是由微管的拉力驱动的,而这种拉力依赖于细胞质动力蛋白。然而,目前尚不清楚动力蛋白如何将中心体带到细胞中心,也就是说,细胞内的哪个结构充当了锚定动力蛋白的底物。在这里,我们提供的证据表明,存在于细胞内细胞器上的一部分动力蛋白,负责细胞器向中心体的运输,它产生了使中心体集中所必需的力。通过使用 C. elegans 全基因组 RNAi 数据库,我们鉴定出 dynein 轻链亚基的一个成员 dyrb-1 可能参与了中心体集中的动力蛋白锚定。DYRB-1 对于细胞器向微管的负端运动是必需的。我们发现中心体集中和细胞器运输的净运动之间存在显著的时间相关性。当 Rab7 和 RILP(它们在哺乳动物细胞中介导细胞器与动力蛋白的结合)被敲低时,中心体集中受到了损害。这些结果表明,沿着微管的细胞器的负端定向运输对于 C. elegans 胚胎中的中心体集中是必需的。基于这一发现,我们提出了一个模型,即沿着微管产生的细胞器运输的反作用力充当了将中心体拉向细胞中心的驱动力。这是我们所知的第一个为细胞质拉力提供中心体集中的力学基础的模型。