RIOK3促进了caspase-10对PAK2的酶解激活

中国生物工程杂志

2012, Vol. 32

Issue (08): 1-8

研究报告

RIOK3促进了caspase-10对PAK2的酶解激活

林颖, 李璞, 单敬轩, 陈晓静, 施慧莉, 霍克克

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

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

LIN Ying, LI Pu, SHAN Jing-xuan, CHEN Xiao-jing, SHI Hui-li, HUO Ke-ke

State Key Laboratory of Genetic Engineering, Institute of Genetics, School of Life Sciences, Fudan University, Shanghai 200433, China

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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。

关键词： RIOK3; PAK2; 酵母双杂交; 细胞凋亡; 酶解激活

Abstract: RIO3, a member of atypical protein kinase, is only discovered in multicellular 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.

Key words: RIOK3 PAK2 Yeast two-hybrid system Cell apoptotic Cleavage-activation

收稿日期: 2012-04-09 出版日期: 2012-08-25

ZTFLH:

Q819

基金资助: 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

通讯作者: 霍克克 E-mail: kkhuo@fudan.edu.cn

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引用本文:

林颖, 李璞, 单敬轩, 陈晓静, 施慧莉, 霍克克. RIOK3促进了caspase-10对PAK2的酶解激活[J]. 中国生物工程杂志, 2012, 32(08): 1-8.

LIN Ying, LI Pu, SHAN Jing-xuan, CHEN Xiao-jing, SHI Hui-li, HUO Ke-ke. RIOK3 Promotes the Cleavage-activation of Caspase-10 on PAK2. China Biotechnology, 2012, 32(08): 1-8.

链接本文:

https://manu60.magtech.com.cn/biotech/CN/ 或 https://manu60.magtech.com.cn/biotech/CN/Y2012/V32/I08/1

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