一种新的中心体复制相关蛋白F46-crp的初步研究
Primary study of a Novel Protein F46-crp That Contributes to Centrosome Duplication
应用一种自身免疫病血清,发现了一种新蛋白F46-crp。检测到其表观分子量为大约60kD,并通过RT-PCR获得了cDNA,制备了单克隆抗体。亚细胞定位研究表明,F46-crp在细胞间期定位于中心体,但有丝分裂开始后消失。以反义RNA沉默F46-crp后,细胞增殖严重受抑并阻抑在S期;反义RNA沉默还导致细胞多中心体和多核化,并最终凋亡。这一研究表明F46-crp是细胞周期相关的蛋白,对于中心体的复制是必要的。
Using a serum from a patient suffering from an autoimmune disease (progressive systemic sclerosis), a novel centrosome related protein F46-crp was detected. We identified the target protein by immunoprecipitation and Western blot and tandem mass spectrometry sequencing. The results show the protein F46-crp has an apparent molecular mass of about 60 kDa. A cDNA of HeLa F46-crp protein was obtained using RT-PCR, and a recombinant F46-crp used to generate monoclonal antibodies. Immunofluorescence microscopy of synchronized HeLa cells labeled with the anti-F46-crp -specific antibodies revealed that F46-crp localized exclusively to the centrosome during interphase, although it completely disappeared at the onset of mitosis. After effectively silencing the F46-crp by antisense RNA, cell growth and proliferation were strongly inhibited and the cell cycle typically stalled at S phase. The silencing also resulted in the formation of poly-centrosomal and multinucleate cells, which finally became apoptotic. These results suggest that F46-crp is a novel centrosome related protein, whose expression is cell cycle-dependent, and it is involved in centrosome duplication.
Shen Enzhi、Jan Marc、魏祎、Wang Yongchao、梁前进、Fan Jinling、Sun Le、Liu Qian
细胞生物学分子生物学
F46-crp,中心体,细胞周期
F46-crp centrosome cell cycle
Shen Enzhi,Jan Marc,魏祎,Wang Yongchao,梁前进,Fan Jinling,Sun Le,Liu Qian.一种新的中心体复制相关蛋白F46-crp的初步研究[EB/OL].(2007-03-09)[2025-08-23].http://www.paper.edu.cn/releasepaper/content/200703-121.点此复制
评论