摘要
hap是从人胚肺细胞系 (WI 38)cDNA文库中克隆的ASY相互作用蛋白基因 ,它为已知基因RTN3的同源体 .hap过表达能引起多种细胞凋亡 .激光共聚焦显微观察显示N端带有EGFP标签的HAP蛋白定位于内质网上 ,C端带有EGFP标签的HAP蛋白则游离分布于整个细胞中 .用Hoechst33342染色观察、DNAladder分析以及流式细胞仪检测均表明 ,定位在内质网上的HAP蛋白高表达的HeLa细胞呈现明显的凋亡特征 ,而游离的HAP高表达的HeLa细胞则没有明显的凋亡现象 .结果表明 ,HAP蛋白的亚细胞定位决定其是否具有诱导细胞凋亡的功能 .
hap, identical to RTN3, was a gene encoding an ASY interacting protein and was identified by cloning complementary DNA from human lung cell line (WI 38) cDNA library using yeast two hybrid screen. It has been verified that overexpression of hap can induce apoptosis in several cells. Confocal microscopy showed HAP protein with EGFP at its N termial located on endoplasmic reticulum(ER) while HAP protein with EGFP at its C terminal suffused in the whole cell. DNA ladder analysis, hoechst33342 staining and flow cytometry analysis demonstrated that apparent apoptosis characteristics exhibited in HeLa cells overexpressing ER locating HAP but not in HeLa cells overexpressing suffused HAP. The subcellular localization decides whether HAP has the function of inducing apoptosis. Combinding with the previous results, it was suggested that HAP was an important part of calcium channel on ER.
出处
《中国生物化学与分子生物学报》
CAS
CSCD
北大核心
2004年第4期528-533,共6页
Chinese Journal of Biochemistry and Molecular Biology
基金
国家自然科学基金 (No .3 0 170 45 5 )资助项目~~
关键词
HAP
内质网
亚细胞定位
凋亡
HAP,endoplasmic reticulum, subcellular localization, apoptosis