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首页> 外文期刊>Journal of Cellular Physiology >Nuclear gamma-tubulin associates with nucleoli and interacts with tumor suppressor protein C53.
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Nuclear gamma-tubulin associates with nucleoli and interacts with tumor suppressor protein C53.

机译：核gamma-tubulin associates和核仁与肿瘤抑制蛋白C53。

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摘要

gamma-Tubulin is assumed to be a typical cytosolic protein necessary for nucleation of microtubules from microtubule organizing centers. Using immunolocalization and cell fractionation techniques in combination with siRNAi and expression of FLAG-tagged constructs, we have obtained evidence that gamma-tubulin is also present in nucleoli of mammalian interphase cells of diverse cellular origins. Immunoelectron microscopy has revealed gamma-tubulin localization outside fibrillar centers where transcription of ribosomal DNA takes place. gamma-Tubulin was associated with nucleolar remnants after nuclear envelope breakdown and could be translocated to nucleoli during mitosis. Pretreatment of cells with leptomycin B did not affect the distribution of nuclear gamma-tubulin, making it unlikely that rapid active transport via nuclear pores participates in the transport of gamma-tubulin into the nucleus. This finding was confirmed by heterokaryon assay and time-lapse imaging of photoconvertible protein Dendra2 tagged to gamma-tubulin. Immunoprecipitation from nuclear extracts combined with mass spectrometry revealed an association of gamma-tubulin with tumor suppressor protein C53 located at multiple subcellular compartments including nucleoli. The notion of an interaction between gamma-tubulin and C53 was corroborated by pull-down and co-immunoprecipitation experiments. Overexpression of gamma-tubulin antagonized the inhibitory effect of C53 on DNA damage G(2) /M checkpoint activation. The combined results indicate that aside from its known role in microtubule nucleation, gamma-tubulin may also have nuclear-specific function(s).
机译：gamma-Tubulin被认为是一个典型的胞质蛋白质所必需的微管的成核从微管组织中心。immunolocalization和细胞分离技术结合siRNAi和表达FLAG-tagged结构,获得的证据表明gamma-tubulin也存在于哺乳动物间期细胞的核仁不同的细胞起源。显微镜显示gamma-tubulin本地化外纤维中心在哪里核糖体DNA的转录。gamma-Tubulin核仁的核膜破裂后残余在有丝分裂可能会转移到核仁。预处理与leptomycin B细胞没有影响核gamma-tubulin的分布,使它不可能快速主动运输通过核孔运输参与gamma-tubulin进入细胞核。证实了异核体试验和延时成像photoconvertible蛋白质gamma-tubulin Dendra2标记。从核中提取免疫沉淀反应结合质谱分析揭示了一个与肿瘤gamma-tubulin协会抑制蛋白质C53位于多个亚细胞的隔间包括核仁。gamma-tubulin之间交互的概念和C53被下拉和证实co-immunoprecipitation实验。过度的gamma-tubulin引起抑制作用对DNA损伤的C53 G (2) / M检查点激活。表明,除了已知的作用微管成核,gamma-tubulin也nuclear-specific函数(年代)。

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