A region in the first exon/intron of rat carnitine palmitoyltransferase Ibeta is involved in enhancement of basal transcription.
Carnitine palmitoyltransferase-Ibeta (CPT-Ibeta) catalyses the transfer of long-chain fatty acids to the enzymes of beta-oxidation of muscle and heart. Transcriptional control of this regulatory protein is relevant to disorders of fatty acid oxidation and the switch to glucose metabolism that occurs in cardiac pathology. The presence of a transcriptional enhancer sequence in the first untranslated exon and first intron of the CPT-Ibeta gene was identified using deletional and mutational analysis, and by ligation of an oleate responsive element (fatty acid response element) to a minimal promoter. The enhancer sequences are contained in the first 40 bases downstream of the transcription start site and increase CPT-Ibeta reporter gene expression independent of any 5' cis-acting elements. Deletion of the first 40 bases of the 3'-untranslated region does not affect the up-regulation of transcription by 10 microM phenylephrine. However, mutation and/or deletion of bases between +11 and +30 dramatically decreases reporter gene expression. Electrophoretic mobility-shift assays reveal two DNA (+11 to +36)-protein complexes that appear cardiac specific. The exon/intron element enhances activation of the heterologous thymidine kinase promoter in a position- and orientation-dependent manner. Therefore we have identified a novel region in the first exon/intron of the CPT-Ibeta gene that acts as a non-classical transcriptional enhancer downstream of regulatory elements characterized previously in the 5'-flanking region of the minimal promoter.
Wang
Guo-Li
GL
Department of Pathology and Laboratory Medicine, University of Texas Medical School at Houston, 6431 Fannin, Houston, TX 77030, U.S.A.
Moore
Meredith L
ML
McMillin
Jeanie B
JB
D020413:3' Untranslated Regions / Q000235:genetics; D000818:Animals; D000831:Animals, Newborn; D001483:Base Sequence; D001665:Binding Sites; D002334:Carnitine O-Palmitoyltransferase / Q000235:genetics*; D002460:Cell Line; D002467:Cell Nucleus / Q000378:metabolism; D004252:DNA Mutational Analysis; D004742:Enhancer Elements, Genetic; D005091:Exons; D005786:Gene Expression Regulation / Q000187:drug effects; D017930:Genes, Reporter; D006801:Humans; D007438:Introns; D007527:Isoenzymes / Q000235:genetics; D007668:Kidney; D008099:Liver / Q000201:enzymology; D008156:Luciferases / Q000235:genetics; D018482:Muscle, Skeletal / Q000201:enzymology; D016297:Mutagenesis, Site-Directed; D009206:Myocardium / Q000201:enzymology* / Q000473:pathology; D019301:Oleic Acid / Q000494:pharmacology; D010084:Oxidation-Reduction; D010656:Phenylephrine / Q000494:pharmacology; D011401:Promoter Regions, Genetic; D051381:Rats; D011994:Recombinant Proteins / Q000378:metabolism; D017384:Sequence Deletion; D014158:Transcription, Genetic / Q000187:drug effects
D020413:3' Untranslated Regions; D007527:Isoenzymes; D011994:Recombinant Proteins; D010656:Phenylephrine; D019301:Oleic Acid; D008156:Luciferases; D002334:Carnitine O-Palmitoyltransferase