›› 2011, Vol. 31 ›› Issue (5): 551-.doi: 10.3969/j.issn.1674-8115.2011.05.006

• Original article (Basic research) • Previous Articles     Next Articles

Transcription factor interactions of TGF-beta signaling pathway identified by yeast two-hybrid technique during mice liver regeneration

XIE Chao1, WANG Fang-yuan2, YUAN Yun-sheng1, GAO Jin3, ZHU Run-zhi4, HAN Wei3, YU Yan1   

  1. 1.Shanghai Key Laboratory of Veterinary Biotechnology, School of Agriculture and Biology, Shanghai Jiaotong University, Shanghai 200240, China;2.Center of Research Laboratory, the International Peace Maternity and Child Health Hospital of China Welfare Institute, Shanghai Jiaotong University School of Medicine, Shanghai 200030, China;3.School of Pharmacy, Shanghai Jiaotong University, Shanghai 200240, China;4.Jiangsu Key Laboratory of Carcinogenesis and Intervention, China Pharmaceutical University, Nanjing 210009, China
  • Online:2011-05-28 Published:2011-05-27
  • Supported by:

    National High-tech R&D Program of China, 2007AA02Z149


Objective To investigate the interactions of four transcription factors of transforming growth factor-β (TGF-β) signaling pathways using yeast two-hybrid technique. Methods CCl4-induced liver injury mouse models were established, and the total RNA of liver tissues were extracted as the template for cDNA by RT-PCR. Activating transcription factor 3 (ATF3), inhibitor of DNA binding 3 (ID3), DNA-damage-inducible transcript 3 (DDIT3) and LIM domains 2 (FHL2), which were differentially expressed detected by liver regeneration gene chip and associated with TGF-β signaling pathway, were served as candidate genes. After gene clone, vector construction and PCR and DNA sequencing, yeast two-hybrid system was used to indentify the interactions among ATF3, ID3, DDIT3 and FHL2 with auto-activation test. Results PCR and DNA sequencing indicated that ATF3, FHL2, DDIT3 and ID3 were successfully cloned, and there was no auto-activation phenomenon after auto-activation test. Seven protein interactions (ID3/ATF3, FHL2/ATF3, ATF3/ATF3, FHL2/FHL2, ID3/FHL2, ID3/ID3 and ID3/DDIT3) were identified by yeast two-hybrid technique. Conclusion Protein interactions among ATF3, FHL2 and ID3 may be one of the possible ways to regulate the TGF-β signaling pathway, and protein-protein interactions may be a key gateway to liver regeneration.

Key words: liver regeneration, transcription factors, yeast two-hybrid system, interactions