CAJ | 학술논문

目的：探讨有丝分裂相关激酶Plk1磷酸化修饰Tara蛋白在调控HeLa细胞胞质分裂及增殖中的作用。方法：运用酵母双杂交、免疫共沉淀、体外沉降实验确定Plk1与Tara两种蛋白是否能够相互结合；采用体内外磷酸化实验明确Plk1对Tara蛋白的磷酸化修饰；采用免疫荧光实验及流式细胞术检测Tara蛋白的磷酸化对HeLa细胞胞质分裂及细胞增殖的影响。结果：Plk1与Tara在体内外均能结合；Tara蛋白在体内外均能够被Plk1磷酸化修饰；过量表达Tara蛋白的磷酸化突变体对HeLa细胞的胞质分裂有明显的抑制作用，细胞增殖明显受阻。结论：Plk1能够与Tara结合，并通过磷酸化修饰Tara蛋白阻碍细胞胞质分裂进程及抑制细胞增殖。
목적：탐토유사분렬상관격매Plk1린산화수식Tara단백재조공HeLa세포포질분렬급증식중적작용。방법：운용효모쌍잡교、면역공침정、체외침강실험학정Plk1여Tara량충단백시부능구상호결합；채용체내외린산화실험명학Plk1대Tara단백적린산화수식；채용면역형광실험급류식세포술검측Tara단백적린산화대HeLa세포포질분렬급세포증식적영향。결과：Plk1여Tara재체내외균능결합；Tara단백재체내외균능구피Plk1린산화수식；과량표체Tara단백적린산화돌변체대HeLa세포적포질분렬유명현적억제작용，세포증식명현수조。결론：Plk1능구여Tara결합，병통과린산화수식Tara단백조애세포포질분렬진정급억제세포증식。
Aim:To investigate the effects of phosphorylation of Tara by Plk1 on cytokinesis and proliferation of cervical cancer HeLa cells.Methods:Using yeast hybridization assay, co-immunoprecipitation and GST pull-down as-say to determine the domain of Plk1 and Tara binding and interaction in vitro and in vivo .To check the phosphoryla-tion of Tara by Plk1 using in vitro phosphorylation assay.Immunoflurescence assay and flow cytometry were used to analyze the effects of phosphorylation of Tara on cytokinesis and proliferation of Hela cells.Results: Plk1 interacted with and phosphorylated Tara in vivo and in vitro .Overexpression of phospho-mimicking mutant of Tara in Hela cells resulted in significant mitosis delay.Conclusion: Plk1-mediated phosphorylation of Tara is essential for cytokinesis and proliferation of HeLa cells.