RIOK3促进了caspase-10对PAK2的酶解激活  

RIOK3 Promotes the Cleavage-activation of Caspase-10 on PAK2

作　　者：林颖[1] 李璞[1] 单敬轩[1] 陈晓静[1] 施慧莉[1] 霍克克[1] 

机构地区：[1]复旦大学生命科学学院遗传学研究所遗传工程国家重点实验室,上海200433

出　　处：《中国生物工程杂志》2012年第8期1-8,共8页China Biotechnology

基　　金：国家高技术研究发展计划(2006AA02A310);国家科技重大专项(2008ZX10003-006;2009ZX09301-011);国家重点基础研究发展计划(2010CB912603)资助项目

摘　　要：RIO3为非典型激酶RIO家族的成员之一,仅在多细胞真核生物中出现,为研究RIOK3蛋白的功能,以其作为诱饵蛋白对成人肝cDNA文库进行酵母双杂交筛选,得到其相互作用蛋白PAK2,并通过细胞内免疫共沉淀和免疫荧光共定位实验验证了该相互作用。实时定量PCR和免疫印迹检测结果显示,RIOK3能够在蛋白水平上降低PAK2表达量。通过CCK-8和细胞凋亡检测,发现二者共表达可以抑制增殖并促进凋亡,并且这一促凋亡效应可以被caspase-10的无酶活最短剪切本caspase-10G所抑制。实验结果显示,RIOK3可能促进了caspase-10对PAK2的酶解,在PAK2的酶解激活途径中发挥重要作用。RIO3, a member of atypical protein kinase, is only discovered in muhicellular eukaryotes. The human liver cDNA library was screened with pDBLeu-RIOK3 as bait plasmid by yeast two-hybrid system and PAK2 was identified as a RIOK3 interactive protein. The interaction was confirmed by co-immunoprecipitation assays and immunofluorescent localization experiments. Results of real-time quantitative PCR and Western blot showed that RIOK3 can reduce the amount of PAK2 at protein expression level. CCK-8 and apoptosis detection experiment showed that co-expression of RIOK3 and PAK2 can inhibit cell proliferation and promote apoptosis, and the apoptotic effect can be inhibited by caspase-10G, which is the minimum isoform of caspase-10 and had no enzyme activity. The experimental results showed that RIOK3 may promote cleavage of caspase-10 on PAK2 and play an important role in the PAK2 cleavage-activation pathway.

关 键 词：RIOK3 PAK2 酵母双杂交 细胞凋亡 酶解激活 

分 类 号：Q819[生物学—生物工程]