›› 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

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Abstract:

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