Using yeast two-hybrid system to identify ECRG2 associated proteins and their possible interactions with ECRG2 gene  

作　　者：Yong-Ping Cui Jian-Bo Wang Xin-Yu Zhang Mei-Xia Bi Li-Ping Guo Shih-Hsin Lu Department of Etiology and Carcinogenesis,Tumor Hospital,Peking Union Medical College & Chinese Academy of Medical Sciences,Beijing 100021,China 

出　　处：《World Journal of Gastroenterology》2003年第9期1892-1896,共5页世界胃肠病学杂志（英文版）

基　　金：G1998051204 the Major State Basic Research Development Program of China,No.G1998051204

摘　　要：AIM: To identify esophageal cancer related gene2 (ECRG2)associated proteins and their possible interactions withECRG2 gene.METHODS: In the yeast forward two-hybrid system, ECRG2was fused with the DNA-binding domain (DBD) of Gal4 andhuman fetal liver cDNA library was fused with thetranscriptional activation domain (AD) of Gal4. We performeda high-stringency scale procedure to screen ECRG2 againsthuman fetal liver cDNA library and characterized positivesby sequence analysis.RESULTS: We found the following 9 putatively associatedproteins. They were metallothionein2A, metallothionein1H,metallothionein1G, ferritin, erythrocyte membrane proteinband4.2, mitochondrial ribosomal protein S12, hypotheticalprotein FLJ10101, and a novel gene whose cDNA was foundto have no strong homology to any other previouslycharacterized gene whose DDBJ/EMBL/GenBank accessionnumber is AF422192 mapped to human chromosome 14q31.CONCLUSION: MT, a potential interaction partner forECRG2, might be involved in the regulation of cellproliferation and apoptosis, and in various physiologicalprocesses. Determination of a reliability score for each singleprotein-protein interaction, especially interaction of ECRG2and MT, permits the assignment of ECRG2 and unannotatedproteins to biological pathways. A further understanding ofthe association between ECRG2 and MT should facilitate the functions of ECRG2 gene.AIM:To identify esophageal cancer related gene2(ECRG2) associated proteins and their possible interactions with ECRG2 gene. METHODS:In the yeast forward two-hybrid system,ECRG2 was fused with the DNA-binding domain(DBD)of Gal4 and human fetal liver cDNA library was fused with the transcriptional activation domain(AD)of Gal4.We performed a high-stringency scale procedure to screen ECRG2 against human fetal liver cDNA library and characterized positives by sequence analysis. RESULTS:We found the following 9 putatively associated proteins.They were metallothionein2A,metallothioneinlH, metallothioneinlG,ferritin,erythrocyte membrane protein band4.2,mitochondrial ribosomal protein S12,hypothetical protein FLJ10101,and a novel gene whose cDNA was found to have no strong homology to any other previously characterized gene whose DDBJ/EMBL/GenBank accession number is AF422192 mapped to human chromosome 14q31. CONCLUSION:MT,a potential interaction partner for ECRG2,might be involved in the regulation of cell proliferation and apoptosis,and in various physiological processes.Determination of a reliability score for each single protein-protein interaction,especially interaction of ECRG2 and MT,permits the assignment of ECRG2 and unannotated proteins to biological pathways.A further understanding of the association between ECRG2 and MT should facilitate the functions of ECRG2 gene.

关 键 词：食道癌 食道癌相关基因2 MT 酵母 

分 类 号：Q78[生物学—分子生物学]