Relative roles of CCAAT/enhancer-binding protein β and cAMP regulatory element-binding protein in controlling transcription of the gene for phosphoenolpyruvate carboxykinase (GTP)

Edwards Park, Austin L. Gurney, Steven E. Nizielski, Parvin Hakimi, Zhodan Cao, Antoon Moorman, Richard W. Hanson

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Abstract

The gene for phosphoenolpyruvate carboxykinase (GTP) (EC 4.1.1.32) (PEPCK) is expressed in a tissuespecific manner in the liver, kidney, and adipose tissue and is regulated by hormones including cAMP and insulin. Previous studies have shown that the CCAAT/ enhancer-binding protein α (C/EBPα) binds to several sites on the PEPCK promoter and activates transcription from the promoter in hepatoma cells. Here, we report that a second member of the C/EBP family, C/ EBPβ, bound to the same sites on the PEPCK promoter. However, C/EBPβ stimulated transcription primarily through the cAMP-responsive element (CRE), which maps between positions -77 to -94, but not at the more 5′-binding sites. In addition, the nuclear factor-1 site, which is immediately adjacent to the CRE in the PEPCK promoter, was also required for the full response of the promoter to cotransfected C/EBPβ. In gel mobility assays, antibodies to both C/EBPβ and the cAMP regulatory element-binding protein (CREB), but not to C/EBPα, "supershifted" DNA-protein complexes formed between a synthetic CRE oligomer and proteins prepared from rat liver nuclei. C/EBPβ mRNA was expressed at low levels in both the periportal and pericentral regions of the liver lobule, whereas expression of the gene for C/EBPα was confined to the pericentral region of the liver lobule. PEPCK gene transcription is greatest in the periportal region of the liver. CREB also bound to the CRE and stimulated transcription of a PEPCK-CAT vector in the presence of an expression vector for the catalytic subunit of protein kinase A. C/ EBPβ and CREB bound to the CRE with similar affinities, both of which were greater than the affinity of C/EBPα. Within 90 min after the administration of dibutyryl cAMP to rats, there was a marked increase in the hepatic concentration of C/EBPβ mRNA and a decrease in the level of mRNA for C/EBPα. These studies indicate that C/EBPβ can regulate PEPCK gene transcription by acting through the CRE and that C/ EBPβ, together with CREB, may contribute to the cAMP responsiveness of the PEPCK promoter.

Original languageEnglish (US)
Pages (from-to)613-619
Number of pages7
JournalJournal of Biological Chemistry
Volume268
Issue number1
StatePublished - Jan 5 1993
Externally publishedYes

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Phosphoenolpyruvate Carboxykinase (GTP)
CCAAT-Enhancer-Binding Proteins
Transcription
Carrier Proteins
Genes
Liver
Messenger RNA
Rats
Cyclic AMP-Dependent Protein Kinase Catalytic Subunits
NFI Transcription Factors

All Science Journal Classification (ASJC) codes

  • Biochemistry

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Relative roles of CCAAT/enhancer-binding protein β and cAMP regulatory element-binding protein in controlling transcription of the gene for phosphoenolpyruvate carboxykinase (GTP). / Park, Edwards; Gurney, Austin L.; Nizielski, Steven E.; Hakimi, Parvin; Cao, Zhodan; Moorman, Antoon; Hanson, Richard W.

In: Journal of Biological Chemistry, Vol. 268, No. 1, 05.01.1993, p. 613-619.

Research output: Contribution to journalArticle

Park, Edwards ; Gurney, Austin L. ; Nizielski, Steven E. ; Hakimi, Parvin ; Cao, Zhodan ; Moorman, Antoon ; Hanson, Richard W. / Relative roles of CCAAT/enhancer-binding protein β and cAMP regulatory element-binding protein in controlling transcription of the gene for phosphoenolpyruvate carboxykinase (GTP). In: Journal of Biological Chemistry. 1993 ; Vol. 268, No. 1. pp. 613-619.
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AU - Nizielski, Steven E.

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