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

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.