Positive and negative regulation of a transfected chimeric tyrosine aminotransferase gene: effect of copy number.
To define a selective system for the study of rat tyrosine aminotransferase (TAT; EC 2.6.1.5) gene expression, we have introduced into cultured cells the selectable bacterial gene gpt linked to TAT gene flanking sequences. After integration in host cell DNA, the chimeric gene exhibits the same pattern of regulation as the TAT gene. In hepatoma cells, its expression is induced after glucocorticoid hormone treatment and repressed after fusion with fibroblasts. In fibroblasts, the chimeric gene is not expressed. The correct pattern of regulation is lost when the number of integrated copies is high. At copy number above 10, the transfected gene responds poorly to glucocorticoids in hepatoma cells. At copy number above 50, the gene is expressed in fibroblasts. Another gene present in the same construction and controlled by the SV40 early promoter and enhancer is positively regulated by glucocorticoids in hepatoma cells but not after fusion with fibroblasts. These data indicate that in hybrid cells, both TAT promoter and glucocorticoid-responsive elements are negatively regulated.
Grange
T
T
Institut Jacques Monod du CNRS, Unité INSERM 257, Université Paris VII, France.
Roux
J
J
Fromont-Racine
M
M
Pictet
R
R
D000818:Animals; D002460:Cell Line; D002678:Chimera; D003907:Dexamethasone / Q000494:pharmacology; D005786:Gene Expression Regulation / Q000187:drug effects; D006822:Hybrid Cells; D051379:Mice; D010430:Pentosyltransferases / Q000235:genetics; D010957:Plasmids; D011401:Promoter Regions, Genetic; D012333:RNA, Messenger / Q000032:analysis / Q000235:genetics; D051381:Rats; D014162:Transfection; D014407:Tumor Cells, Cultured; D014444:Tyrosine Transaminase / Q000235:genetics*
D012333:RNA, Messenger; D003907:Dexamethasone; D010430:Pentosyltransferases; C024827:xanthine phosphoribosyltransferase; D014444:Tyrosine Transaminase