Tanenbaum Marvin E, Galjart Niels, van Vugt Marcel A T M, Medema René H
Division of Molecular Biology, Netherlands Cancer Institute, Amsterdam, The Netherlands
EMBO J. 2006 Jan 11;25(1):45-57. doi: 10.1038/sj.emboj.7600916. Epub 2005 Dec 15.
CLIP-170 is a microtubule 'plus end tracking' protein involved in several microtubule-dependent processes in interphase. At the onset of mitosis, CLIP-170 localizes to kinetochores, but at metaphase, it is no longer detectable at kinetochores. Although RNA interference (RNAi) experiments have suggested an essential role for CLIP-170 during mitosis, the molecular function of CLIP-170 in mitosis has not yet been revealed. Here, we used a combination of high-resolution microscopy and RNAi-mediated depletion to study the function of CLIP-170 in mitosis. We found that CLIP-170 dynamically localizes to the outer most part of unattached kinetochores and to the ends of growing microtubules. In addition, we provide evidence that a pool of CLIP-170 is transported along kinetochore-microtubules by the dynein/dynactin complex. Interference with CLIP-170 expression results in defective chromosome congression and diminished kinetochore-microtubule attachments, but does not detectibly affect microtubule dynamics or kinetochore-microtubule stability. Taken together, our results indicate that CLIP-170 facilitates the formation of kinetochore-microtubule attachments, possibly through direct capture of microtubules at the kinetochore.
CLIP-170是一种微管“正端追踪”蛋白,参与间期的多种微管依赖过程。在有丝分裂开始时,CLIP-170定位于动粒,但在中期,动粒上不再能检测到它。尽管RNA干扰(RNAi)实验表明CLIP-170在有丝分裂过程中起重要作用,但其在有丝分裂中的分子功能尚未揭示。在这里,我们结合高分辨率显微镜和RNAi介导的耗竭来研究CLIP-170在有丝分裂中的功能。我们发现CLIP-170动态定位于未附着动粒的最外层部分以及生长中的微管末端。此外,我们提供证据表明,CLIP-170的一部分由动力蛋白/动力蛋白激活蛋白复合物沿动粒微管运输。干扰CLIP-170的表达会导致染色体排列异常和动粒微管附着减少,但不会明显影响微管动力学或动粒微管稳定性。综上所述,我们的结果表明CLIP-170可能通过在动粒处直接捕获微管来促进动粒微管附着的形成。