生长抑制蛋白4核定位信号和PHD指状结构域共同调控丝裂原活化蛋白激酶通路中ERK1的激活  

Nuclear Localization Signal and PHD Domain of ING4 Play an Important Role in Stimulation of ERK1 MAPK

作　　者：潘春阳[1] 王克生 王勤婉 韩泽广[1] 张新[1] 

机构地区：[1]华东理工大学生物工程学院,上海市200237 [2]国家人类基因组南方研究中心,上海市201203

出　　处：《医学分子生物学杂志》2008年第2期100-104,共5页Journal of Medical Molecular Biology

基　　金：国家高技术研究发展计划(863计划)(No.2006AA02A305)~~

摘　　要：目的探索生长抑制蛋白4(inhibitor of growth 4,ING4)的核定位信号(nuclear localization sig-nal,NLS)和PHD指状结构域(plant homeodomain)在刺激丝裂原活化蛋白激酶(MAPK)细胞信号通路成员ERK1磷酸化中的作用。方法通过构建ING4的NLS和PHD结构域突变或缺失重组质粒,将上述质粒分别与ERK1表达载体共转染HEK-293细胞,Western印迹检测ERK1总蛋白和磷酸化ERK1蛋白水平。结果在HEK-293细胞中,ING4蛋白分别缺失NLS和PHD结构域后,其对MAPK细胞信号通路成员ERK1磷酸化的促进作用均降低,突变ING4的NLS结构域中两个重要位点Lys-Lys(KK)和Arg-Ala-Arg-Ser-Lys(RARSK)并不改变对ERK1激活的能力,但突变ING4 PHD结构域中RKKK位点则改变了调控ERK1磷酸化的能力。结论ING4的NLS和PHD指状结构域同时参与了刺激MAPK细胞信号通路成员ERK1的激活。Objective In order to identify the domain of ING4 involved in the modulation of ERK1 MAPK pathway. Methods The plasmids of ING4 site mutants or plasmids with deletion of its NLS and PHD domain were constructed. ING4 mutants or part-deletion vectors were co-transfected by introducing ERKl-expressing vector into HEK-293 cells and the phosphorylation of ERK1 was detected by Western blotting. Results The stimulation of ING4 to ERK1 was abrogated with the deletion of its NLS or PHD domain. Furthermore, RKKK site in PHD domain is also important for the modulation of ERK1 phosphorylation. Conclusion Both nuclear localization signal and PHD domain are important for ING4 regulation of ERK1 phosphorylation.

关 键 词：生长抑制蛋白4 核定位信号 PHD指状结构域 ERK1 

分 类 号：Q71[生物学—分子生物学]