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Using yeast two-hybrid system to identify ECRG2 associated proteins and their possible interactions with ECRG2 gene
Yong-Ping Cui1, Jian-Bo Wang, Xin-Yu Zhang
1Department of Etiology and Carcinogenesis, Tumor Hospital, Peking Union Medical College and Chinese Academy of Medical Sciences, Beijing 100021, China.
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, metallothionein1H, metallothionein1G, 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.