Morrison E E, Wardleworth B N, Askham J M, Markham A F, Meredith D M
Molecular Medicine Unit, University of Leeds, St. James's University Hospital, UK.
Oncogene. 1998 Dec 31;17(26):3471-7. doi: 10.1038/sj.onc.1202247.
The characteristics of the adenomatous polyposis coli (APC) associated protein EB1 were examined in mammalian cells. By immunocytochemistry EB1 was shown to be closely associated with the microtubule cytoskeleton throughout the cell cycle. In interphase cells EB1 was associated with microtubules along their full length but was often particularly concentrated at their tips. During early mitosis, EB1 was localized to separating centrosomes and associated microtubules, while at metaphase it was associated with the spindle poles and associated microtubules. During cytokinesis EB1 was strongly associated with the midbody microtubules. Treatment with nocodazole caused a diffuse redistribution of EB1 immunoreactivity, whereas treatment with cytochalasin D had no effect. Interestingly, treatment with taxol abolished the EB1 association with microtubules. In nocodazole washout experiments EB1 rapidly became associated with the centrosome and repolymerizing microtubules. In taxol wash-out experiments EB1 rapidly re-associated with the microtubule cytoskeleton, resembling untreated control cells within 10 min. Immunostaining of SW480 cells, which contain truncated APC incapable of interaction with EB1, showed that the association of EB1 with microtubules throughout the cell cycle was not dependent upon an interaction with APC. These results suggest a role for EB1 in the control of microtubule dynamics in mammalian cells.
在哺乳动物细胞中研究了腺瘤性结肠息肉病相关蛋白EB1的特性。通过免疫细胞化学方法发现,在整个细胞周期中EB1都与微管细胞骨架紧密相关。在间期细胞中,EB1与微管的全长相关联,但通常特别集中在微管的末端。在有丝分裂早期,EB1定位于分离的中心体和相关的微管上,而在中期它与纺锤体极和相关的微管相关联。在胞质分裂期间,EB1与中体微管强烈相关。用诺考达唑处理导致EB1免疫反应性的弥散性重新分布,而用细胞松弛素D处理则没有效果。有趣的是,用紫杉醇处理消除了EB1与微管的关联。在诺考达唑洗脱实验中,EB1迅速与中心体和重新聚合的微管相关联。在紫杉醇洗脱实验中,EB1迅速重新与微管细胞骨架相关联,在10分钟内类似于未处理的对照细胞。对含有截短的APC且无法与EB1相互作用的SW480细胞进行免疫染色,结果表明EB1在整个细胞周期中与微管的关联并不依赖于与APC的相互作用。这些结果表明EB1在哺乳动物细胞微管动力学控制中发挥作用。