两种存活蛋白变异剪接体在HeLa细胞中的表达定位及相互作用  

Expression and Location of Two Survivin Splice Variants and Their Interaction in HeLa Cells

作　　者：邵红伟[1] 张文峰[1] 胡青莲[1] 黄树林[1] 

机构地区：[1]广东药学院生命科学与生物制药学院生物制药研究所,广州510006

出　　处：《中国生物化学与分子生物学报》2009年第12期1131-1136,共6页Chinese Journal of Biochemistry and Molecular Biology

基　　金："重大新药创制"科技重大专项"十一五"计划项目(No.2009ZX09103-708);广东省高校优秀青年创新人才培育项目(No.2008342);广东药学院师资队伍建设经费(No.43555014)资助~~

摘　　要：存活蛋白(survivin)是重要的肿瘤相关抗原基因,在肿瘤的发生发展中起着重要的作用.除了标准的剪接形式外,它至少还编码2种变异剪接产物—存活蛋白-2B和存活蛋白-ΔEx3,这2个变异剪接体所编码的蛋白具有不同的生物学功能.为研究这2个变异剪接体在肿瘤细胞中的相互作用情况,本实验利用增强型青色荧光蛋白(enhanced cyan fluorescent protein,ECFP)和增强型黄色荧光蛋白(enhanced yellowfluorescent protein,EYFP)分别标记存活蛋白-2B和存活蛋白-ΔEx3.首先通过激光共聚焦扫描显微镜观察它们的细胞定位;同时利用荧光共振能量转移(fluorescence resonanceenergytransfer,FRET)技术研究两者在细胞内的相互作用情况.研究结果表明,存活蛋白-2B主要分布在细胞质中,而存活蛋白-ΔEx3则主要分布在细胞核内,少量分布在细胞质中;FRET分析结果显示,两者仅在细胞质中存在着很弱的相互作用,表明两者很可能是通过某种间接的方式发挥功能上的相互调节作用.本研究为进一步探讨存活蛋白变异剪接体的生物学功能及相互作用机制奠定了基础.Survivin is one of tumor associated antigen genes that plays an important role in cancer development.Survivin-2B and survivin-ΔEx3 are the two normal splice variants（isoforms） encoding two different proteins.To investigate the interaction between the two isoforms,survivin-2B was labeled with enhanced cyan fluorescent protein（ECFP） and survivin-ΔEx3 was labeled with enhanced yellow fluorescent protein（EYFP）.The expression and subcellular localization of survivin-2B and survivin-ΔEx3 were observed with a confocal laser scanning microscope（CLSM）,and their intermolecular interaction was quantified by fluorescence resonance energy transfer（FRET） method.It was shown that survivin-2B expressed in the cytoplasm,but most of the survivin-ΔEx3 localized in the nucleus.Very weak intermolecular interaction between the two isoforms was observed in the cytoplasm.These results might provide useful materials for further clarifying the biological function and the interactions of survivin-2B and survivin-ΔEx3.

关 键 词：存活蛋白-2B 存活蛋白-ΔEx3 荧光共振能量转移 相互作用 

分 类 号：Q291[生物学—细胞生物学]