Ambrose J Christian, Shoji Tsubasa, Kotzer Amanda M, Pighin Jamie A, Wasteneys Geoffrey O
University of British Columbia, Vancouver, British Columbia, Canada.
Plant Cell. 2007 Sep;19(9):2763-75. doi: 10.1105/tpc.107.053777. Epub 2007 Sep 14.
Controlling microtubule dynamics and spatial organization is a fundamental requirement of eukaryotic cell function. Members of the ORBIT/MAST/CLASP family of microtubule-associated proteins associate with the plus ends of microtubules, where they promote the addition of tubulin subunits into attached kinetochore fibers during mitosis and stabilize microtubules in the vicinity of the plasma membrane during interphase. To date, nothing is known about their function in plants. Here, we show that the Arabidopsis thaliana CLASP protein is a microtubule-associated protein that is involved in both cell division and cell expansion. Green fluorescent protein-CLASP localizes along the full length of microtubules and shows enrichment at growing plus ends. Our analysis suggests that CLASP promotes microtubule stability. clasp-1 T-DNA insertion mutants are hypersensitive to microtubule-destabilizing drugs and exhibit more sparsely populated, yet well ordered, root cortical microtubule arrays. Overexpression of CLASP promotes microtubule bundles that are resistant to depolymerization with oryzalin. Furthermore, clasp-1 mutants have aberrant microtubule preprophase bands, mitotic spindles, and phragmoplasts, indicating a role for At CLASP in stabilizing mitotic arrays. clasp-1 plants are dwarf, have significantly reduced cell numbers in the root division zone, and have defects in directional cell expansion. We discuss possible mechanisms of CLASP function in higher plants.
控制微管动力学和空间组织是真核细胞功能的基本要求。微管相关蛋白的ORBIT/MAST/CLASP家族成员与微管的正端结合,在有丝分裂期间,它们促进微管蛋白亚基添加到附着的动粒纤维中,并在间期稳定质膜附近的微管。迄今为止,关于它们在植物中的功能尚不清楚。在这里,我们表明拟南芥CLASP蛋白是一种微管相关蛋白,参与细胞分裂和细胞扩张。绿色荧光蛋白-CLASP定位于微管的全长,并在生长的正端富集。我们的分析表明CLASP促进微管稳定性。clasp-1 T-DNA插入突变体对微管去稳定药物高度敏感,并表现出根皮层微管阵列分布更稀疏但排列有序。CLASP的过表达促进了对oryzalin解聚有抗性的微管束。此外,clasp-1突变体具有异常的微管前期带、有丝分裂纺锤体和成膜体,表明At CLASP在稳定有丝分裂阵列中起作用。clasp-1植物矮小,根分裂区的细胞数量显著减少,并且在定向细胞扩张方面存在缺陷。我们讨论了CLASP在高等植物中发挥功能的可能机制。