Yeast one-hybrid system used to identify the binding proteins for rat glutathione S-transferase P enhancer I

Biomed Environ Sci. 2002 Mar;15(1):36-40.

Abstract

Objective: To detect the trans-factors specifically binding to the strong enhancer element (GPEI) in the upstream of rat glutathione S-transferase P (GST-P) gene.

Methods: Yeast one-hybrid system was used to screen rat lung MATCHMAKER cDNA library to identify potential trans-factors that can interact with core sequence of GPEI(cGPEI). Electrophoresis mobility shift assay (EMSA) was used to analyze the binding of transfactors to cGPEI.

Results: cDNA fragments coding for the C-terminal part of the transcription factor c-Jun and rat adenine nucleotide translocator (ANT) were isolated. The binding of c-Jun and ANT to GPEI core sequence were confirmed.

Conclusions: Rat c-jun transcriptional factor and ANT may interact with cGPEI. They could play an important role in the induced expression of GST-P gene.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Biological Assay / methods
  • Carrier Proteins / chemistry
  • Carrier Proteins / isolation & purification
  • DNA Primers
  • DNA, Complementary / genetics*
  • Enhancer Elements, Genetic / genetics*
  • Enzyme Induction
  • Gene Expression Regulation*
  • Gene Library
  • Glutathione Transferase / genetics*
  • Lung
  • Rats
  • Sequence Analysis, DNA
  • Yeasts

Substances

  • Carrier Proteins
  • DNA Primers
  • DNA, Complementary
  • Glutathione Transferase