Identification of glucocorticoid-induced genes in rat hepatoma cells by isolation of cloned cDNA sequences.
The expression of specific cellular genes in M1.19 rat hepatoma cells involves glucocorticoid regulation by mechanisms that are not well understood. To approach this problem we cloned cDNA prepared from dexamethasone-induced poly(A)-RNA and used a comparative colony hybridization method to identify recombinant clones containing hormone-regulated sequences. Two such cDNA clones, p1394 and p255, hybridize to a homogeneous RNA species of 900 nucleotides that is present in high abundance in 24-hr-induced cells but is undetectable in uninduced cells. This RNA can be seen as early as 1 hr after dexamethasone stimulation. Inhibition of protein synthesis with cycloheximide significantly reduces the accumulation of the RNA but does not abolish the induction response. In normal adult rat liver the RNA is abundant, and this RNA is induced by dexamethasone in adrenalectomized rats. Plasmids p1394 and p255 contain sequences that are homologous to the mRNA coding for the acute-phase reactant protein alpha 1-acid glycoprotein. Two other cDNA clones, p655 and p333, hybridize to a more heterogeneous RNA species 200-400 nucleotides in size with a lower induction response to dexamethasone. Southern blot analysis of M1.19 genomic DNA indicates that p1394 and p255 are complementary to a single DNA fragment, whereas p655 and p333 are complementary to repetitive sequences in the M1.19 genome. It appears that the genetic domain of glucocorticoid control in M1.19 rat hepatoma cells involves low copy number genes such as alpha 1-acid glycoprotein as well as repetitive sequence elements.
D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D003001:Cloning, Molecular; D004247:DNA / Q000032:analysis*; D004262:DNA Restriction Enzymes; D004273:DNA, Neoplasm / Q000235:genetics; D003907:Dexamethasone / Q000494:pharmacology*; D005796:Genes / Q000187:drug effects*; D008114:Liver Neoplasms, Experimental / Q000235:genetics*; D009693:Nucleic Acid Hybridization; D010957:Plasmids; D011061:Poly A / Q000235:genetics; D012313:RNA / Q000235:genetics; D012333:RNA, Messenger; D012334:RNA, Neoplasm / Q000235:genetics; D051381:Rats
D004273:DNA, Neoplasm; D012333:RNA, Messenger; D012334:RNA, Neoplasm; D011061:Poly A; D012313:RNA; D003907:Dexamethasone; D004247:DNA; D004262:DNA Restriction Enzymes