S-pombe CLASP needs dynein, not EB1 or CLIP170, to induce microtubule instability and slows polymerization rates at cell tips in a dynein-dependent manner
S-pombe CLASP needs dynein, not EB1 or CLIP170, to induce microtubule instability and slows polymerization rates at cell tips in a dynein-dependent manner
复制标题
DOI：
10.1101/gad.381306
复制
发表时间：
2006-09-01
影响因子：
10.5
通讯作者：
Hagan, Iain M.
中科院分区：
文献类型：
--
作者：
Grallert, Agnes;Beuter, Christoph;Hagan, Iain M.
粟酒裂殖酵母CLIP 170相关蛋白（CLASP）Peg1在筛选纺锤体形成缺陷的突变体和筛选拮抗EB1功能的分子中被鉴定。有条件的peg1.1突变体使我们能够识别Peg1功能的关键特征。首先，Peg1需要形成纺锤体和星形微管，但不稳定的间期微管。第二，Peg1需要减慢间期微管的聚合速率，这些微管在细胞尖端与皮质建立端对接触。第三，Peg1拮抗S. pombe CLIP170（Tip1）和EB1（Mal3）。第四，虽然Peg1类似于更高的真核CLASPs的物理关联与Mal3和Tip1，无论是Tip1或Mal3是必要的Peg1不稳定间期微管或它与微管。相反，Mal3和Tip1都不需要Peg1与微管或细胞尖端相关联。一致地，虽然mal3.Delta和tip1.Delta破坏线性生长，但破坏peg1（+）没有。第五，peg1.1表型类似于那些由缺失单个重链或两个轻链的裂殖酵母动力蛋白。此外，PEG 1+操纵产生的所有间期表型依赖于动力蛋白功能。因此，S.粟酒裂殖酵母CLASP对间期微管行为的影响比EB 1或CLIP 170更接近于动力蛋白。
The Schizosaccharomyces pombe CLIP170-associated protein (CLASP) Peg1 was identified in a screen for mutants with spindle formation defects and a screen for molecules that antagonized EB1 function. The conditional peg1.1 mutant enabled us to identify key features of Peg1 function. First, Peg1 was required to form a spindle and astral microtubules, yet destabilized interphase microtubules. Second, Peg1 was required to slow the polymerization rate of interphase microtubules that establish end-on contact with the cortex at cell tips. Third, Peg1 antagonized the action of S. pombe CLIP170 (Tip1) and EB1 (Mal3). Fourth, although Peg1 resembled higher eukaryotic CLASPs by physically associating with both Mal3 and Tip1, neither Tip1 nor Mal3 was required for Peg1 to destabilize interphase microtubules or for it to associate with microtubules. Conversely, neither Mal3 nor Tip1 required Peg1 to associate with microtubules or cell tips. Consistently, while mal3.Delta and tip1.Delta disrupted linear growth, corrupting peg1(+) did not. Fifth, peg1.1 phenotypes resembled those arising from deletion of the single heavy or both light chains of fission yeast dynein. Furthermore, all interphase phenotypes arising from peg1+ manipulation relied on dynein function. Thus, the impact of S. pombe CLASP on interphase microtubule behavior is more closely aligned to dynein than EB1 or CLIP170.