Transcriptional and post-transcriptional mechanisms can regulate cell-specific expression of the human Pi-class glutathione S-transferase gene.
Previous studies from this laboratory have identified transcriptional mechanisms that are utilized to increase expression of the human glutathione S-transferase gene GSTP1 in a multidrug-resistant derivative (VCREMS) of the human mammary carcinoma cell line MCF7 [Moffat, McLaren and Wolf (1994) J. Biol. Chem. 269, 16397-16402]. The data presented here provide strong evidence that post-transcriptional mechanisms can also play an important role in determining cell-specific expression of the GSTP1 gene. GSTP1 mRNA levels were shown to be elevated 3.1-fold in the human bladder carcinoma cell line EJ compared with VCREMS cells. Despite this observation, transient transfection assays revealed a decreased rate of GSTP1 promoter activity in EJ cells. Indeed, GSTP1 transcriptional repressor activity, mediated by a region located between nucleotides -105 and -86 (as we have previously described in MCF7 cells), was observed in EJ cells. However, in contrast with our results in MCF7 cells, the EJ repressor activity did not displace the essential nuclear complex bound to the C1 promoter element (-73 to -54) in vitro. In addition, competition experiments indicated that an AP-1-like protein is an integral component of the C1-bound complex in EJ cells. Interestingly, experiments utilizing actinomycin D to inhibit transcription demonstrated significantly greater stability of GSTP1 mRNA in EJ cells than in VCREMS cells. These findings suggest that cell-specific differences in the rates of GSTP1 mRNA decay provide the predominant mechanism responsible for elevated expression of the GSTP1 gene in EJ cells.
Moffat
G J
GJ
Imperial Cancer Research Fund, Molecular Pharmacology Unit, Biomedical Research Centre, Ninewells Hospital and Medical School, Dundee DD1 9SY, Scotland, U.K.
McLaren
A W
AW
Wolf
C R
CR
D001483:Base Sequence; D001943:Breast Neoplasms; D002460:Cell Line; D002467:Cell Nucleus / Q000378:metabolism; D003609:Dactinomycin / Q000494:pharmacology; D005260:Female; D015971:Gene Expression Regulation, Enzymologic; D005982:Glutathione Transferase / Q000096:biosynthesis*; D006801:Humans; D007527:Isoenzymes / Q000096:biosynthesis; D007700:Kinetics; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D015345:Oligonucleotide Probes; D009928:Organ Specificity; D011401:Promoter Regions, Genetic; D012323:RNA Processing, Post-Transcriptional; D012333:RNA, Messenger / Q000378:metabolism; D011994:Recombinant Proteins / Q000096:biosynthesis; D017384:Sequence Deletion; D014158:Transcription, Genetic / Q000187:drug effects; D014407:Tumor Cells, Cultured; D001749:Urinary Bladder Neoplasms
D007527:Isoenzymes; D015345:Oligonucleotide Probes; D012333:RNA, Messenger; D011994:Recombinant Proteins; D003609:Dactinomycin; D005982:Glutathione Transferase