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Regulation of phosphoenolpyruvate carboxykinase gene transcription by insulin and cAMP: reciprocal actions on initiation and elongation.

Abstract
Nuclei isolated from H4IIE rat hepatoma cells were used in an in vitro run-on assay, with probes directed against various regions of the phosphoenolpyruvate carboxykinase [GTP: oxaloacetate carboxy-lyase (transphosphorylating); EC 4.1.1.32] gene, to analyze whether transcription proceeds uniformly across this gene in response to insulin and cAMP treatment. Fewer polymerase II complexes were associated with the phosphoenolpyruvate carboxykinase gene after insulin treatment, as compared with cAMP-treated cells, but they were distributed uniformly, so insulin does not block transcription at a discrete site, nor does it cause gradual, but progressive, premature termination. The phosphoenolpyruvate carboxykinase primary transcript was synthesized at a rate of about 2500 nucleotides per min in cAMP-treated cells and about 1000 nucleotides per min in insulin-treated cells. Thus insulin retards transcript elongation in comparison with cAMP, but this action does not account for the total effect insulin has on transcription. After insulin treatment, few, if any, nascent transcripts are associated with the first 69 nucleotides of the gene, whereas in cAMP-treated cells the opposite is true. These observations lead us to suggest that both insulin and cAMP exert their primary effects directly at the level of transcription initiation, but in opposite ways.
AuthorsK Sasaki, D K Granner
JournalProceedings of the National Academy of Sciences of the United States of America (Proc Natl Acad Sci U S A) Vol. 85 Issue 9 Pg. 2954-8 (May 1988) ISSN: 0027-8424 [Print] United States
PMID2834722 (Publication Type: Journal Article, Research Support, U.S. Gov't, P.H.S.)
Chemical References
  • Insulin
  • Cyclic AMP
  • Phosphoenolpyruvate Carboxykinase (GTP)
Topics
  • Animals
  • Cell Line
  • Cyclic AMP (pharmacology)
  • Insulin (pharmacology)
  • Liver Neoplasms, Experimental (enzymology, genetics)
  • Peptide Chain Elongation, Translational (drug effects)
  • Peptide Chain Initiation, Translational (drug effects)
  • Phosphoenolpyruvate Carboxykinase (GTP) (genetics)
  • Rats
  • Transcription, Genetic (drug effects)

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