D816V突变型KIT特异性诱导COS-1非洲绿猴肾细胞核不均一核糖核蛋白 L(HNRNPL)和HNRNPK磷酸化  

D816V mutation of KIT specifically induces phosphorylation of HNRNPL and HNRNPK in COS-1 cells

作　　者：张良颖 张少婷 范朝阳 蒋宗英 李淑璟 刘安步 孙建民 ZHANG Liangying;ZHANG Shaoting;FAN Zhaoyang;JIANG Zongying;LI Shujing;LIU Anbu;SUN Jianmin(School of Basic Medical Science,Ningxia Medical University,NHC Key Laboratory of Metabolic Cardiovascular Diseases Research,Medical Science and Technology Research Center,Yinchuan 750004,China)

机构地区：[1]宁夏医科大学基础医学院,国家卫生健康委代谢性心血管疾病研究重点实验室,医学科学技术研究中心,宁夏银川750004

出　　处：《细胞与分子免疫学杂志》2023年第2期138-143,共6页Chinese Journal of Cellular and Molecular Immunology

基　　金：国家自然科学基金(81660473)。

摘　　要：目的探究Ⅲ类酪氨酸激酶受体KIT D816V突变对核不均一核糖核蛋白L(HNRNPL)和HNRNPK的调控作用。方法在COS-1非洲绿猴肾细胞表达野生型KIT或KIT D816V,或与HNRNPL或HNRNPK共表达,用免疫沉淀法和Western blot法检测KIT活化及HNRNPL、HNRNPK的磷酸化,通过激光共聚焦显微镜检测KIT、HNRNPL、HNRNPK在COS-1细胞中的定位。结果野生型KIT需要其配基干细胞因子(SCF)的刺激发生活化,而KIT D816V无需SCF刺激即可发生自活化。此外,KIT D816V可诱导HNRNPL和HNRNPK发生磷酸化,而野生型KIT无此功能。HNRNPL和HNRNPK在细胞核表达,野生型KIT在细胞膜和细胞质表达,而KIT D816V主要表达于细胞质。结论野生型KIT活化需要其配体SCF参与,而KIT D816V无需SCF刺激即可发生自活化,并能特异性诱导HNRNPL和HNRNPK磷酸化。Objectivee To study the regulation of D816V mutation of Ⅲ tyrosine kinase receptor KIT on RNA binding proteins HNRNPL and HNRNPK.Methods In COS-1 cells,wild-type KIT or KIT D816V mutation were expressed alone or together with HNRNPL or HNRNPK.Activation of KIT and phosphorylation of HNRNPL and HNRNPK were detected by immunoprecipitation and Western blot analysis.The localization of KIT,HNRNPL and HNRNPK in COS-1 cells were examined by confocal microscopy.Results Wild-type KIT needs to bind its ligand stem cell factor(SCF)for phosphorylation,while KIT D816V could auto-phosphorylation without SCF stimulation.In addition,KIT D816V can induce phosphorylation of HNRNPL and HNRNPK,which is not possible in wild-type KIT.HNRNPL and HNRNPK are expressed in the nucleus,and wild-type KIT is expressed in cytosol and cell membrane,while KIT D816V is mainly found in cytosol.Conclusion Wild-type KIT needs SCF binding for activation,while KIT D816V can autoactivate without SCF stimulation,and induces phosphorylation of HNRNPL and HNRNPK specifically.

关 键 词：KIT D816V 核不均一核糖核蛋白L(HNRNPL) 核不均一核糖核蛋白K(HNRNPK) 

分 类 号：R733.7[医药卫生—肿瘤] R733.73[医药卫生—临床医学] R392-33