Mps1 phosphorylation sites regulate the function of centrin 2 in centriole assembly.
Mps1 phosphorylation sites regulate the function of centrin 2 in centriole assembly.
复制标题
MPS1磷酸化位点调节中心蛋白2在中心组装中的功能。
DOI：
10.1091/mbc.e10-04-0298
复制
发表时间：
2010-12
影响因子：
3.3
通讯作者：
Fisk HA
中科院分区：
文献类型：
--
作者：
Yang CH;Kasbek C;Majumder S;Yusof AM;Fisk HA
我们发现，虽然Centrin 2是certain的中心粒组装，它是一个Mps 1基板，刺激典型的和异常的中心粒组装由两种不同的Mps 1依赖的机制，HsSas-6依赖和独立。中心蛋白2磷酸化也是Mps 1驱动成熟中心粒产生的能力所必需的。不可降解的Mps 1 Δ12/13蛋白驱动中心粒的过度产生，这表明Mps 1磷酸化中心体蛋白的一个子集来驱动新中心粒的组装。在这里，我们确定了三个Mps 1磷酸化位点内的中心粒蛋白中心蛋白2（Cetn 2）。尽管中心粒可以在Cetn 2不存在的情况下组装，但中心粒组装在Cetn 2不存在的情况下减弱。虽然野生型Cetn 2可以补偿这种衰减，但不可磷酸化的版本不能。此外，过度表达Cetn 2会导致Mps 1依赖性中心粒过度产生，这需要Cetn 2内的三个Mps 1磷酸化位点中的每一个，并且通过模拟这些位点中的任何一个的磷酸化而大大加剧。野生型Cetn 2产生过量的病灶，这些病灶在HsSas-6耗尽的细胞中能够作为有丝分裂纺锤体极，这表明Cetn 2可以独立于侧手翻组织中心粒蛋白的子集。然而，由磷酸化模拟Cetn 2突变体引起的中心粒过度产生需要HsSas-6，这表明Cetn 2磷酸化刺激经典的中心粒组装途径。此外，在缺乏Cetn 2的情况下，Mps 1 Δ12/13不能驱动能够募集γ-微管蛋白的成熟中心粒的产生，并且不可磷酸化的Cetn 2突变体不能补偿这种缺陷并加剧Cetn 2消耗。总之，我们的数据表明，Mps 1依赖性磷酸化Cetn 2刺激经典的中心粒组装途径。
We show that while Centrin2 is dispensable for centriole assembly, it is an Mps1 substrate that stimulates canonical and aberrant centriole assembly by two different Mps1-dependent mechanisms, HsSas-6–dependent and –independent. Centrin2 phosphorylation is also required for the ability of Mps1 to drive production of mature centrioles. The nondegradable Mps1Δ12/13 protein drives centriole overproduction, suggesting that Mps1 phosphorylates a subset of centrosomal proteins to drive the assembly of new centrioles. Here we identify three Mps1 phosphorylation sites within the centriolar protein Centrin 2 (Cetn2). Although centrioles can be assembled in the absence of Cetn2, centriole assembly is attenuated in the absence of Cetn2. While wild-type Cetn2 can compensate for this attenuation, a nonphosphorylatable version cannot. In addition, overexpressing Cetn2 causes Mps1-dependent centriole overproduction that requires each of the three Mps1 phosphorylation sites within Cetn2 and is greatly exacerbated by mimicking phosphorylation at any of these sites. Wild-type Cetn2 generates excess foci that are competent as mitotic spindle poles in HsSas-6–depleted cells, suggesting that Cetn2 can organize a subset of centriolar proteins independently of cartwheels. However, centriole overproduction caused by a phosphomimetic Cetn2 mutant requires HsSas-6, suggesting that Cetn2 phosphorylation stimulates the canonical centriole assembly pathway. Moreover, in the absence of Cetn2, Mps1Δ12/13 cannot drive the production of mature centrioles capable of recruiting γ-Tubulin, and a nonphosphorylatable Cetn2 mutant cannot compensate for this defect and exacerbates Cetn2 depletion. Together, our data suggest that Mps1-dependent phosphorylation of Cetn2 stimulates the canonical centriole assembly pathway.