Louie Ryan K, Bahmanyar Shirin, Siemers Kathleen A, Votin Violet, Chang Paul, Stearns Tim, Nelson W James, Barth Angela I M
Department of Molecular and Cellular Physiology, Stanford University School of Medicine, Stanford, CA 94305, USA.
J Cell Sci. 2004 Mar 1;117(Pt 7):1117-28. doi: 10.1242/jcs.00939. Epub 2004 Feb 17.
Adenomatous polyposis coli (APC) and End-binding protein 1 (EB1) localize to centrosomes independently of cytoplasmic microtubules (MTs) and purify with centrosomes from mammalian cell lines. Localization of EB1 to centrosomes is independent of its MT binding domain and is mediated by its C-terminus. Both APC and EB1 preferentially localize to the mother centriole and EB1 forms a cap at the end of the mother centriole that contains the subdistal appendages as defined by epsilon-tubulin localization. Like endogenous APC and EB1, fluorescent protein fusions of APC and EB1 localize preferentially to the mother centriole. Depletion of EB1 by RNA interference reduces MT minus-end anchoring at centrosomes and delays MT regrowth from centrosomes. In summary, our data indicate that APC and EB1 are functional components of mammalian centrosomes and that EB1 is important for anchoring cytoplasmic MT minus ends to the subdistal appendages of the mother centriole.
腺瘤性结肠息肉病蛋白(APC)和末端结合蛋白1(EB1)定位于中心体,不依赖于细胞质微管(MTs),并可从哺乳动物细胞系的中心体中纯化出来。EB1定位于中心体与其微管结合结构域无关,而是由其C末端介导。APC和EB1都优先定位于母中心粒,EB1在母中心粒末端形成一个帽,该帽包含由ε-微管蛋白定位所定义的亚远端附属物。与内源性APC和EB1一样,APC和EB1的荧光蛋白融合物优先定位于母中心粒。通过RNA干扰耗尽EB1会减少微管负端在中心体的锚定,并延迟微管从中心体的重新生长。总之,我们的数据表明,APC和EB1是哺乳动物中心体的功能成分,并且EB1对于将细胞质微管负端锚定到母中心粒的亚远端附属物很重要。