The centrosomal protein C-Nap1 is required for cell cycle-regulated centrosome cohesion.
The centrosomal protein C-Nap1 is required for cell cycle-regulated centrosome cohesion.
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DOI：
10.1083/jcb.151.4.837
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发表时间：
2000-11-13
期刊：
影响因子：
--
通讯作者：
Nigg EA
中科院分区：
文献类型：
--
作者：
Mayor T;Stierhof YD;Tanaka K;Fry AM;Nigg EA
复制中心体在分裂间期配对，但在有丝分裂开始时分离。虽然控制中心体凝聚和分离的机制对整个细胞周期的中心体功能很重要，但它们仍然知之甚少。最近，我们提出，C-Nap 1，一种新的中心体蛋白，是一个结构的一部分，连接父母的中心粒在细胞周期调控的方式。为了测试这个模型，我们对C-Nap 1进行了详细的结构-功能分析。我们证明，抗体介导的干扰C-Nap 1功能的原因中心体分裂，无论细胞周期阶段。分裂发生在亲本中心粒之间，并且不依赖于完整微管或微丝网络的存在。中心体分裂也可以通过截短的C-Nap 1突变体的过表达来诱导，但不是全长蛋白。针对C-Nap 1的不同结构域的抗体证明，这种蛋白质在有丝分裂期间从纺锤体两极解离，但在细胞分裂结束时在中心体重新积聚。在免疫电镜中使用相同的抗体显示，C-Nap 1被限制在中心粒的近端结构域，表明推定的接头结构必须包含额外的蛋白质。我们的结论是，C-Nap 1是一个动态的，细胞周期调节的结构，介导中心粒-中心粒凝聚力的关键组成部分。
Duplicating centrosomes are paired during interphase, but are separated at the onset of mitosis. Although the mechanisms controlling centrosome cohesion and separation are important for centrosome function throughout the cell cycle, they remain poorly understood. Recently, we have proposed that C-Nap1, a novel centrosomal protein, is part of a structure linking parental centrioles in a cell cycle–regulated manner. To test this model, we have performed a detailed structure–function analysis on C-Nap1. We demonstrate that antibody-mediated interference with C-Nap1 function causes centrosome splitting, regardless of the cell cycle phase. Splitting occurs between parental centrioles and is not dependent on the presence of an intact microtubule or microfilament network. Centrosome splitting can also be induced by overexpression of truncated C-Nap1 mutants, but not full-length protein. Antibodies raised against different domains of C-Nap1 prove that this protein dissociates from spindle poles during mitosis, but reaccumulates at centrosomes at the end of cell division. Use of the same antibodies in immunoelectron microscopy shows that C-Nap1 is confined to the proximal end domains of centrioles, indicating that a putative linker structure must contain additional proteins. We conclude that C-Nap1 is a key component of a dynamic, cell cycle–regulated structure that mediates centriole–centriole cohesion.
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