Differential potency and trans-activation of normal and mutant T24 human H-ras1 gene promoters.
We have employed a short-term transfection assay system in which we monitored the transient expression of the chloramphenicol acetyltransferase (CAT) gene linked to the promoter region of the normal and mutant T24 H-ras1 gene or the human epsilon-globin gene in Chinese hamster lung (CHL) cells or cells derived from them which carry and express one or the other of the polyoma virus early genes. Our findings can be summarized as follows: (i) The mutant T24 H-ras1 promoter region behaves as a stronger promoter than the H-ras1 gene in all these types of cells as well as in rat 208F fibroblast cells. (ii) In CHL cells expressing the polyoma large T antigen the normal and mutant T24 Ha-ras1 promoters are not trans-activated in these cells and only a 2.5-fold activation of the epsilon-globin promoter is observed. (iii) In cells expressing the polyoma middle T antigen both the normal and mutant H-ras1 are trans-activated whereas transcription from the epsilon-globin promoter is not affected when compared to the normal CHL cells. (iv) In cells expressing the polyoma small T antigen the normal and mutant H-ras1 as well as the epsilon-globin promoters are trans-activated. We suggest from these data that a tissue-specific element exists in the promoter region of the H-ras1 gene and that the polyoma middle and small T antigens trigger the expression of proteins that trans-activate these promoters.
Spandidos
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DA
Beatson Institute for Cancer Research, Bearsden, Glasgow, Scotland.
Pintzas
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A
D000123:Acetyltransferases / Q000235:genetics; D000818:Animals; D000952:Antigens, Polyomavirus Transforming / Q000235:genetics; D002460:Cell Line; D015500:Chloramphenicol O-Acetyltransferase; D006224:Cricetinae; D004274:DNA, Recombinant; D005786:Gene Expression Regulation; D005914:Globins / Q000235:genetics; D006801:Humans; D009154:Mutation; D010957:Plasmids; D011401:Promoter Regions, Genetic; D011518:Proto-Oncogene Proteins / Q000235:genetics*; D016283:Proto-Oncogene Proteins p21(ras); D014162:Transfection
D000952:Antigens, Polyomavirus Transforming; D004274:DNA, Recombinant; D011518:Proto-Oncogene Proteins; D005914:Globins; D000123:Acetyltransferases; D015500:Chloramphenicol O-Acetyltransferase; C501009:HRAS protein, human; D016283:Proto-Oncogene Proteins p21(ras)