Rapid insulin-stimulated accumulation of an mRNA encoding a proline-rich protein.
By differential hybridization screening of a cDNA library derived from insulin-stimulated cells, we selected a clone which hybridized to an mRNA species that rapidly accumulated in response to insulin. The insert from this clone encoded a putative polypeptide of Mr 33,600, pI 11.2; because the protein was enriched in proline residues (14.4 mol %) and contained three Pro-Pro-Pro-Pro repeats, we have tentatively labeled it tris-tetraprolin (TTP). The function of this protein is not known, but it contains two regions very rich in proline (30-40 mol %); similar proline-rich regions have been shown to be involved in transcriptional activation by other proteins. The mRNA (2.0 kilobases) encoding the TTP protein was essentially undetectable in serum-deprived HIR 3.5 cells, but accumulated dramatically within 10 min of stimulation by insulin. This effect appeared to be due to insulin acting through the intrinsic protein-tyrosine kinase activity of its own receptor. Insulin induction of TTP mRNA accumulation was prevented by actinomycin D and superinduced by cycloheximide. Accumulation of TTP mRNA was also stimulated by a variety of growth factors and active phorbol esters; however, the insulin effect was virtually normal in cells depleted of protein kinase C. A single TTP gene appeared to be present in the mouse genome. This gene joins the group of genes whose members are rapidly transcribed in response to insulin and other mitogens.
Howard Hughes Medical Institute Laboratories, Durham, North Carolina 27710.
W S
WS
Lai
D J
DJ
Stumpo
P J
PJ
Blackshear
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D015723:Gene Library; D007328:Insulin / Q000494:pharmacology*; D051379:Mice; D008969:Molecular Sequence Data; D015345:Oligonucleotide Probes; D010455:Peptides / Q000235:genetics*; D010750:Phosphoproteins / Q000235:genetics; D011392:Proline; D055232:Proline-Rich Protein Domains; D014176:Protein Biosynthesis; D011487:Protein Conformation; D011493:Protein Kinase C / Q000037:antagonists & inhibitors; D012333:RNA, Messenger / Q000187:drug effects / Q000235:genetics* / Q000378:metabolism; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D014158:Transcription, Genetic / Q000187:drug effects*
D007328:Insulin; D015345:Oligonucleotide Probes; D010455:Peptides; D010750:Phosphoproteins; D012333:RNA, Messenger; D011392:Proline; D011493:Protein Kinase C; D013755:Tetradecanoylphorbol Acetate