Marceiller Jérôme, Drechou Anne, Durand Geneviève, Perez Franck, Poüs Christian
Laboratoire de Biochimie et Biologie Cellulaire, EAD 1595, Faculté de Pharmacie, 5 rue J.B. Clément, 92296 Châtenay-Malabry cedex, France.
Exp Cell Res. 2005 Apr 1;304(2):483-92. doi: 10.1016/j.yexcr.2004.12.007. Epub 2005 Jan 11.
Upon recovery from nocodazole treatment, microtubules from cultured epithelial cells exhibit unusual properties: they re-grow as fast as any highly dynamic microtubule, but they are also protected against disassembly when challenged with nocodazole like the stable microtubules of steady-state cells. Exploring the mechanism that underlies this protection, we found that it was sensitive to ATP treatment and that it involved conventional kinesin. Kinesin localized at the growing end or along nascent microtubules. Its inhibition using a dominant-negative construct for cargo binding, or by micro-injecting an anti-kinesin heavy chain antibody that impairs motor activity, resulted in the partial or total loss of microtubule protection. Finally, in an ex vivo elongation assay, we found that kinesin also participates in the control of microtubule re-growth. Altogether, our findings suggest that kinesin is involved in an early microtubule protection process that is linked to the control of their dynamics during their early growth phase.
从诺考达唑处理中恢复后,培养的上皮细胞中的微管表现出不同寻常的特性:它们重新生长的速度与任何高度动态的微管一样快,但当受到诺考达唑挑战时,它们也像稳态细胞的稳定微管一样受到保护而不被拆卸。探索这种保护作用背后的机制时,我们发现它对ATP处理敏感,并且涉及传统驱动蛋白。驱动蛋白定位于生长末端或新生微管上。使用用于货物结合的显性负性构建体抑制它,或者通过显微注射损害运动活性的抗驱动蛋白重链抗体抑制它,会导致微管保护作用部分或完全丧失。最后,在体外延伸试验中,我们发现驱动蛋白也参与微管重新生长的控制。总之,我们的研究结果表明,驱动蛋白参与了早期微管保护过程,该过程与微管在早期生长阶段的动态控制有关。