Dayanandan Rejith, Butler Rachel, Gordon-Weeks Phillip R, Matus Andrew, Kaech Stefanie, Lovestone Simon, Anderton Brian H, Gallo Jean-Marc
Department of Neuroscience, Institute of Psychiatry, King's College London, United Kingdom.
Cell Motil Cytoskeleton. 2003 Mar;54(3):237-47. doi: 10.1002/cm.10096.
The adenomatous polyposis coli (APC) tumour suppressor protein is a component of the Wnt signalling pathway in which it plays a major role in controlling nuclear accumulation of beta-catenin and hence in the modulation of beta-catenin-regulated gene transcription. APC also associates with microtubules at the ends of cytoplasmic extensions in epithelial cells, a distribution that can be reproduced in COS cells ectopically expressing APC. To examine the effect of APC on microtubule properties, we monitored directly the behaviour of APC and of APC-decorated microtubules by time-lapse imaging of cytoplasmic extensions in live COS cells expressing APC tagged with a green fluorescent protein. On the proximal part of microtubules, APC was visualised as particulate material moving unidirectionally towards the plus end of microtubules. The distal parts of microtubules were uniformly decorated by APC and were animated by a motile behaviour in the form of aperiodic bending. This behaviour is likely to be the consequence of compression forces acting on microtubules encountering obstacles while elongating. The majority of APC-decorated microtubules in transfected COS cells was sensitive to depolymerisation by nocodazole, but they contained detyrosinated and acetylated alpha-tubulin, suggesting a reduction in the rate of subunit exchange at their growing end. Taken together, these results demonstrate that microtubule domains uniformly decorated by APC display dynamic and motile properties that may be significant for the postulated role of APC in targeting microtubules to specialised membrane sites.
腺瘤性结肠息肉病蛋白(APC)是Wnt信号通路的一个组成部分,在该通路中,它在控制β-连环蛋白的核内积累方面发挥主要作用,从而调节β-连环蛋白调控的基因转录。APC还在上皮细胞胞质延伸末端与微管结合,这种分布可在异位表达APC的COS细胞中重现。为了研究APC对微管特性的影响,我们通过对表达绿色荧光蛋白标记的APC的活COS细胞的胞质延伸进行延时成像,直接监测了APC以及APC修饰的微管的行为。在微管的近端部分,APC表现为颗粒物质单向朝着微管的正端移动。微管的远端部分被APC均匀修饰,并以非周期性弯曲的运动行为活跃起来。这种行为可能是微管在伸长时遇到障碍物而受到压缩力的结果。转染的COS细胞中大多数被APC修饰的微管对诺考达唑介导的解聚敏感,但它们含有去酪氨酸化和乙酰化的α-微管蛋白,这表明其生长末端的亚基交换速率降低。综上所述,这些结果表明,被APC均匀修饰的微管结构域具有动态和运动特性,这可能对APC将微管靶向特定膜位点的假定作用具有重要意义。