Identification and partial characterization of genes that are transactivated by different pathways in quiescent mouse cells stimulated with serum.
We have identified four cDNA clones, cl-1, cl-5, cl-15, and cl-16, that represent genes induced by serum in resting mouse 3T3 cells. Partial sequence analysis of the four cDNAs indicated that cl-15 corresponds to the mouse beta-actin gene. Comparison of the DNA sequences of the other three clones with the sequence data bank (Genbank) showed little homology to other known DNA sequences and thus represent novel genes. The level of the mRNAs corresponding to the four genes began to increase in resting cells following serum stimulation, reached a peak between 5 h and 8 h and then started to decline. Inhibitors of transcription diminished the induction of the mRNAs corresponding to the four genes. Cycloheximide and anisomycin had little effect on the induction of beta actin mRNA while the induction of the other three genes was suppressed by the same inhibitors. 12-O-Tetradecanoylphorbol-13-acetate and the calcium ionophore A23187 enhanced the expression of the cl-16 mRNA while epidermal growth factor, fibroblast growth factor, or insulin enhanced the expression of cl-1- and cl-5-specific transcripts. The level of beta-actin mRNA was elevated in resting cells by epidermal growth factor and 12-O-tetradecanoylphorbol-13-acetate and to a lesser extent by fibroblast growth factor, insulin, and dibutyryl cyclic AMP-elevating agents. Pertussis toxin, an inhibitor of the action of G proteins, did not significantly suppress the activation of the four genes by serum. However, 2-aminopurine, a protein kinase inhibitor, suppressed the induction of the four transcripts in serum-stimulated cells. The possible pathways involved in the activation of these genes in resting cells are discussed.
Boeggeman
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Department of Microbiology, Georgetown University School of Medicine, Washington, D.C. 20007.
Masibay
A S
AS
Qasba
P K
PK
Sreevalsan
T
T
D000199:Actins / Q000235:genetics; D000818:Animals; D002453:Cell Cycle / Q000235:genetics*; D002478:Cells, Cultured; D003001:Cloning, Molecular; D005786:Gene Expression Regulation; D015723:Gene Library; D006133:Growth Substances / Q000502:physiology; D051379:Mice; D011061:Poly A / Q000096:biosynthesis; D014176:Protein Biosynthesis / Q000187:drug effects; D012333:RNA, Messenger / Q000096:biosynthesis; D014158:Transcription, Genetic / Q000187:drug effects; D015533:Transcriptional Activation
D000199:Actins; D006133:Growth Substances; D012333:RNA, Messenger; D011061:Poly A