Nuclear Factor I Regulates Expression of the Gene for Phosphoenolpyruvate Carboxykinase (GTP)

Deborah R Crawford,Patrick Leahy,Ching Y Hu,Ali Chaudhry,Richard Gronostajski,Gregory Grossman,Jason Woods,Parvin Hakimi,William J Roesler,Richard W Hanson

doi:10.1074/jbc.273.22.13387

Abstract

Nuclear factor-I (NFI) binds to the phosphoenolpyruvate carboxykinase (GTP) (PEPCK) gene promoter immediately 5' to the cAMP regulatory element (CRE). This suggests an interaction between NFI and factors that bind the CRE. Of the four NFI isoforms expressed in mammalian tissues, NFI-A and -B stimulate basal transcription from the PEPCK gene promoter in HepG2 cells, while NFI-C and -X are slightly inhibitory. All four NFI isoforms abrogate the 20-fold protein kinase Ac (PKAc)-mediated induction of transcription from the PEPCK gene promoter. Normal PKAc-mediated induction was noted when the CRE was moved 10 base pairs 3' of its original location. However if the CRE was moved 5 base pairs 3', placing it out of phase with the other elements in the promoter, or moved 5' to -285 (the P3(I) site in the promoter), some PKA-mediated stimulation was lost. The NFI-C isoform effectively inhibited PKAc induction regardless of the relative positions of the CRE and the NFI binding sites. NFI-C also abrogated cAMP regulatory element-binding protein (CREB)-induced activity of wild type and mutant PEPCK promoters. There was some cooperativity in the binding of CREB and NFI to their respective binding sites but this did not appear to be physiologically important.

Highlights

Phosphoenolpyruvate carboxykinase (GTP) (EC 4.1.1.32) (PEPCK)1 is a key enzyme in hepatic and renal gluconeogenesis, whose activity is tightly regulated at the transcriptional level
The proximity of the P1 and CRE sites in the PEPCK gene promoter raises the possibility of interactions between the NFI isoforms and protein(s) that bind the CRE
Footprint analysis of the PEPCK gene promoter has shown that both the P1 and CRE sites on the endogenous PEPCK gene were occupied under conditions of basal gene expression [13, 14]

Summary

Introduction

Phosphoenolpyruvate carboxykinase (GTP) (EC 4.1.1.32) (PEPCK)1 is a key enzyme in hepatic and renal gluconeogenesis, whose activity is tightly regulated at the transcriptional level. The proximity of the P1 and CRE sites in the PEPCK gene promoter raises the possibility of interactions between the NFI isoforms and protein(s) that bind the CRE.

Results

Conclusion