Cloning and characterization of the promoter for the liver isoform of the rat carnitine palmitoyltransferase I (L-CPT I) gene

Park, A. Edwards; Steffen, L. Michelle; Song, Shulan; Park, M. Vicki; Cook, Atlanta G.

doi:10.1042/bj3300217

Cited by 42 publications

(51 citation statements)

References 46 publications

(67 reference statements)

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“…Alignment of these alternative ends with the complete gene sequence that has since been published has now confirmed this. As in the case of the mammalian CPT1A (35,36) and CPT1B promoters (37)(38)(39)(40), those for DmCPT1 are TATAless. Both promoter regions of the D. melanogaster CPT1 contain a number of sequence motifs with a high degree of similarity to known transcription factor-binding sites.…”

Section: Tablementioning

confidence: 99%

Alternative Exon Usage in the Single CPT1 Gene of Drosophila Generates Functional Diversity in the Kinetic Properties of the Enzyme

Price

Jackson

Müller

et al. 2010

Journal of Biological Chemistry

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The Drosophila melanogaster genome contains only one CPT1 gene (Jackson, V. N., Cameron, J. M., Zammit, V. A., and Price, N. T. (1999) Biochem. J. 341, 483-489). We have now extended our original observation to all insect genomes that have been sequenced, suggesting that a single CPT1 gene is a universal feature of insect genomes. We hypothesized that insects may be able to generate kinetically distinct variants by alternative splicing of their single CPT1 gene. Analysis of the insect genomes revealed that (a) the single CPT1 gene in each and every insect genome contains two alternative exons and (ii) in all cases, the putative alternative splicing site occurs within a small region corresponding to 21 amino acid residues that are known to be essential for the binding of substrates and of malonyl-CoA in mammalian CPT1A. We performed PCR analyses of mRNA from different Drosophila tissues; both of the anticipated splice variants of CPT1 mRNA were found to be expressed in all of the tissues tested (both in larvae and adults), with the expression level for one of the splice variants being significantly different between flight muscle and the fat body of adult Drosophila. Heterologous expression of the full-length cDNAs corresponding to the two putative variants of Drosophila CPT1 in the yeast Pichia pastoris revealed two important differences between the properties of the two variants: (i) their affinity (K 0.5 ) for one of the substrates, palmitoyl-CoA, differed by 5-fold, and (ii) the sensitivity to inhibition by malonyl-CoA at fixed, higher palmitoyl-CoA concentrations was 2-fold different and associated with different kinetics of inhibition. These data indicate that alternative splicing that specifically affects a structurally crucial region of the protein is an important mechanism through which functional diversity of CPT1 kinetics is generated from the single gene that occurs in insects.Lipid metabolism in insects has largely been investigated from the perspective of energy supply to flight muscle, particularly in migratory flight. Because their lipid metabolism is less complex than in vertebrates, insects are considered a good model system in which to study the fundamental aspects of fat metabolism. The increasing availability of insect gene sequence data and completely sequenced genomes allows comparative analyses with mammalian data and suggests that many of the fundamental molecular mechanisms involved in fatty acid metabolism and their regulation are conserved. Although in most insects there are major differences in the glycerolipids used to transport lipid between tissues (diacylglycerol in insects versus triacylglycerol in vertebrates), the transfer of the long chain acyl moiety across the mitochondrial inner membrane is likely to be substantially similar to that in vertebrates and to involve the carnitine shuttle (1). We have previously shown that the fruit fly Drosophila melanogaster has only a single CPT1 (carnitine palmitoyltransferase 1) gene (2). When we expressed the full-length cDNA (Berkeley Dro...

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Section: Tablementioning

confidence: 99%

Alternative Exon Usage in the Single CPT1 Gene of Drosophila Generates Functional Diversity in the Kinetic Properties of the Enzyme

Price

Jackson

Müller

et al. 2010

Journal of Biological Chemistry

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“…6). In these experiments, we used primary rat hepatocytes because we had cloned the rat CPT-I␣ gene (9). Rat hepatocytes were exposed to T3 for 24 h or left in serum-free media.…”

Section: Fig 4 Amino-terminal Amino Acids Of Pgc-1␣ Stimulate Basalmentioning

confidence: 99%

“…Real Time PCR-RNA was extracted from primary rat hepatocytes using RNA-Stat-60 (Tel-Test) as described previously (9). The mRNA abundance was measured by real time PCR.…”

Section: Transient Transfection Of Luciferasementioning

confidence: 99%

Peroxisomal Proliferator-activated Receptor-γ Coactivator-1α (PGC-1α) Enhances the Thyroid Hormone Induction of Carnitine Palmitoyltransferase I (CPT-Iα)

Zhang

Song

et al. 2004

Journal of Biological Chemistry

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Carnitine palmitoyltransferase I (CPT-I) catalyzes the rate-controlling step in the pathway of mitochondrial fatty acid oxidation. Thyroid hormone will stimulate the expression of the liver isoform of CPT-I (CPT-I␣). This induction of CPT-I␣ gene expression requires the thyroid hormone response element in the promoter and sequences within the first intron. The peroxisomal proliferator-activated receptor-␥ coactivator-1␣ (PGC-1␣) is a coactivator that promotes mitochondrial biogenesis, mitochondrial fatty acid oxidation, and hepatic gluconeogenesis. In addition, PGC-1␣ will stimulate the expression of CPT-I␣ in primary rat hepatocytes. Here we report that thyroid hormone will increase PGC-1␣ mRNA and protein levels in rat hepatocytes. In addition, overexpression of PGC-1␣ will enhance the thyroid hormone induction of CPT-I␣ indicating that PGC-1␣ is a coactivator for thyroid hormone. By using chromatin immunoprecipitation assays, we show that PGC-1␣ is associated with both the thyroid hormone response element in the CPT-I␣ gene promoter and the first intron of the CPT-I␣ gene. Our data demonstrate that PGC-1␣ participates in the stimulation of CPT-I␣ gene expression by thyroid hormone and suggest that PGC-1␣ is a coactivator for thyroid hormone.

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“…Previously, we cloned the promoter of the CPT-I␣ gene (21). The CPT-I␣ promoter is TATA-less, and basal expression is driven by Sp1 and NF-Y (22).…”

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confidence: 99%

Thyroid Hormone Regulates Carnitine Palmitoyltransferase Iα Gene Expression through Elements in the Promoter and First Intron

Jansen¹,

Cook

Song

et al. 2000

Journal of Biological Chemistry

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Cloning and characterization of the promoter for the liver isoform of the rat carnitine palmitoyltransferase I (L-CPT I) gene

Cited by 42 publications

References 46 publications

Alternative Exon Usage in the Single CPT1 Gene of Drosophila Generates Functional Diversity in the Kinetic Properties of the Enzyme

Alternative Exon Usage in the Single CPT1 Gene of Drosophila Generates Functional Diversity in the Kinetic Properties of the Enzyme

Peroxisomal Proliferator-activated Receptor-γ Coactivator-1α (PGC-1α) Enhances the Thyroid Hormone Induction of Carnitine Palmitoyltransferase I (CPT-Iα)

Thyroid Hormone Regulates Carnitine Palmitoyltransferase Iα Gene Expression through Elements in the Promoter and First Intron

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