Identification of basal and cyclic AMP regulatory elements in the promoter of the phosphoenolpyruvate carboxykinase gene

Journal Article · Mon Aug 01 04:00:00 EDT 1988 · Mol. Cell. Biol.; (United States)

DOI:https://doi.org/10.1128/MCB.8.8.3467· OSTI ID:7095919

Quinn, P G; Wong, T W; Magnuson, M A; Shabb, J B; Granner, D K

Promoter elements important for basal and cyclic AMP (cAMP)-regulated expression of the phosphoenolpyruvate carboxykinase (PEPCK) gene have been identified by analysis of a series of PEPCK promoter mutations in transfection experiments. Fusion genes containing wild-type and mutated PEPCK promoter sequences from -600 to +69 base pairs (bp) fused to the coding sequence for chloramphenicol acetyltransferase were studied. Internal deletion mutations that replaced specific bases with a 10-bp linker within the region from - 129 bp to - 18 bp of the PEPCK promoter were examined. In addition, wild-type and mutated DNA templates were used as probes in DNase I protection experiments to determine sites of protein-DNA interaction. The PEPCK promoter contains a binding site for nuclear factor 1-CAAT. Deletion of the 5' end of this binding site reduced the size of the DNase I footprint in this region but had no effect on promoter activity. In contrast, deletion or disruption of the 3' end of this binding site completely eliminated protein binding and reduced promoter activity by 50%. Deletion of core sequences of the cAMP regulatory element (CRE) resulted in loss of cAMP responsiveness and an 85% decrease in basal promoter activity, indicating that the CRE also functions as a basal stimulatory element.

• Research Organization: Dept. of Molecular Physiology and Biophysics, Vanderbilt Univ., Nashville, TN (US)
• OSTI ID: 7095919
• Journal Information: Mol. Cell. Biol.; (United States), Journal Name: Mol. Cell. Biol.; (United States) Vol. 8:8; ISSN MCEBD
• Country of Publication: United States
• Language: English

Related Subjects

550200* -- Biochemistry
59 BASIC BIOLOGICAL SCIENCES
AMP
CARBOXYLIC ACIDS
CYCLASES
DATA
DNA SEQUENCING
DNA-ASE
ENZYME ACTIVITY
ENZYMES
ESTERASES
EXPERIMENTAL DATA
GENE MUTATIONS
GENE OPERONS
GENE REGULATION
GENES
HYDROLASES
INFORMATION
KETO ACIDS
LYASES
MUTATIONS
NUCLEOTIDES
NUMERICAL DATA
ORGANIC ACIDS
ORGANIC COMPOUNDS
PHENOTYPE
PHOSPHODIESTERASES
PHOSPHORUS-GROUP TRANSFERASES
PHOSPHOTRANSFERASES
PYRUVIC ACID
STRUCTURAL CHEMICAL ANALYSIS
TRANSFERASES