Yeast one-hybrid system used to identify the binding proteins for rat glutathione S-transferase P enhancer I.
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.
Liao
Ming-Xiang
MX
National Laboratory of Medical Molecular Biology, Institute of Basic Medical Sciences, Institute of Laboratory Animal Science, Chinese Academy of Medical Sciences, Peking Union Medical College, Beijing 100005, China.
Liu
Dong-Yuan
DY
Zuo
Jin
J
Fang
Fu-De
FD
D000818:Animals; D001681:Biological Assay / Q000379:methods; D002352:Carrier Proteins / Q000737:chemistry / Q000302:isolation & purification; D017931:DNA Primers; D018076:DNA, Complementary / Q000235:genetics*; D004742:Enhancer Elements, Genetic / Q000235:genetics*; D004790:Enzyme Induction; D005786:Gene Expression Regulation; D015723:Gene Library; D005982:Glutathione Transferase / Q000235:genetics*; D008168:Lung; D051381:Rats; D017422:Sequence Analysis, DNA; D015003:Yeasts
D002352:Carrier Proteins; D017931:DNA Primers; D018076:DNA, Complementary; D005982:Glutathione Transferase