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. 1999 Jun 1;340(Pt 2):425–432.

Expression of the rat liver carnitine palmitoyltransferase I (CPT-Ialpha) gene is regulated by Sp1 and nuclear factor Y: chromosomal localization and promoter characterization.

M L Steffen 1, W R Harrison 1, F F Elder 1, G A Cook 1, E A Park 1
PMCID: PMC1220267  PMID: 10333485

Abstract

Carnitine palmitoyltransferase (CPT)-I catalyses the transfer of long-chain fatty acids from CoA to carnitine for translocation across the mitochondrial inner membrane. Expression of the 'liver' isoform of the CPT-I gene (CPT-Ialpha) is subject to developmental, hormonal and tissue-specific regulation. To understand the basis for control of CPT-Ialpha gene expression, we have characterized the proximal promoter of the CPT-Ialpha gene. Here, we report the sequence of 6839 base pairs of the promoter and the localization of the rat CPT-Ialpha gene to region q43 on chromosome 1. Our studies show that the first 200 base pairs of the promoter are sufficient to drive transcription of the CPT-Ialpha gene. Within this region are two sites that bind both Sp1 and Sp3 transcription factors. In addition, nuclear factor Y (NF-Y) binds the proximal promoter. Mutation at the Sp1 or NF-Y sites severely decreases transcription from the CPT-Ialpha promoter. Other protein binding sites were identified within the first 200 base pairs of the promoter by DNase I footprinting, and these elements contribute to CPT-Ialpha gene expression. Our studies demonstrate that CPT-Ialpha is a TATA-less gene which utilizes NF-Y and Sp proteins to drive basal expression.

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Selected References

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  1. Bigger C. B., Melnikova I. N., Gardner P. D. Sp1 and Sp3 regulate expression of the neuronal nicotinic acetylcholine receptor beta4 subunit gene. J Biol Chem. 1997 Oct 10;272(41):25976–25982. doi: 10.1074/jbc.272.41.25976. [DOI] [PubMed] [Google Scholar]
  2. Britton C. H., Mackey D. W., Esser V., Foster D. W., Burns D. K., Yarnall D. P., Froguel P., McGarry J. D. Fine chromosome mapping of the genes for human liver and muscle carnitine palmitoyltransferase I (CPT1A and CPT1B). Genomics. 1997 Feb 15;40(1):209–211. doi: 10.1006/geno.1996.4539. [DOI] [PubMed] [Google Scholar]
  3. Britton C. H., Schultz R. A., Zhang B., Esser V., Foster D. W., McGarry J. D. Human liver mitochondrial carnitine palmitoyltransferase I: characterization of its cDNA and chromosomal localization and partial analysis of the gene. Proc Natl Acad Sci U S A. 1995 Mar 14;92(6):1984–1988. doi: 10.1073/pnas.92.6.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Brown N. F., Weis B. C., Husti J. E., Foster D. W., McGarry J. D. Mitochondrial carnitine palmitoyltransferase I isoform switching in the developing rat heart. J Biol Chem. 1995 Apr 14;270(15):8952–8957. doi: 10.1074/jbc.270.15.8952. [DOI] [PubMed] [Google Scholar]
  5. Chatelain F., Kohl C., Esser V., McGarry J. D., Girard J., Pegorier J. P. Cyclic AMP and fatty acids increase carnitine palmitoyltransferase I gene transcription in cultured fetal rat hepatocytes. Eur J Biochem. 1996 Feb 1;235(3):789–798. doi: 10.1111/j.1432-1033.1996.00789.x. [DOI] [PubMed] [Google Scholar]
  6. Chen T. T., Wu R. L., Castro-Munozledo F., Sun T. T. Regulation of K3 keratin gene transcription by Sp1 and AP-2 in differentiating rabbit corneal epithelial cells. Mol Cell Biol. 1997 Jun;17(6):3056–3064. doi: 10.1128/mcb.17.6.3056. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Choi S. J., Oh D. H., Song C. S., Roy A. K., Chatterjee B. Molecular cloning and sequence analysis of the rat liver carnitine octanoyltransferase cDNA, its natural gene and the gene promoter. Biochim Biophys Acta. 1995 Nov 7;1264(2):215–222. doi: 10.1016/0167-4781(95)00146-8. [DOI] [PubMed] [Google Scholar]
  8. Cole T. J., Blendy J. A., Monaghan A. P., Krieglstein K., Schmid W., Aguzzi A., Fantuzzi G., Hummler E., Unsicker K., Schütz G. Targeted disruption of the glucocorticoid receptor gene blocks adrenergic chromaffin cell development and severely retards lung maturation. Genes Dev. 1995 Jul 1;9(13):1608–1621. doi: 10.1101/gad.9.13.1608. [DOI] [PubMed] [Google Scholar]
  9. Esser V., Britton C. H., Weis B. C., Foster D. W., McGarry J. D. Cloning, sequencing, and expression of a cDNA encoding rat liver carnitine palmitoyltransferase I. Direct evidence that a single polypeptide is involved in inhibitor interaction and catalytic function. J Biol Chem. 1993 Mar 15;268(8):5817–5822. [PubMed] [Google Scholar]
  10. Esser V., Brown N. F., Cowan A. T., Foster D. W., McGarry J. D. Expression of a cDNA isolated from rat brown adipose tissue and heart identifies the product as the muscle isoform of carnitine palmitoyltransferase I (M-CPT I). M-CPT I is the predominant CPT I isoform expressed in both white (epididymal) and brown adipocytes. J Biol Chem. 1996 Mar 22;271(12):6972–6977. doi: 10.1074/jbc.271.12.6972. [DOI] [PubMed] [Google Scholar]
  11. Gorski K., Carneiro M., Schibler U. Tissue-specific in vitro transcription from the mouse albumin promoter. Cell. 1986 Dec 5;47(5):767–776. doi: 10.1016/0092-8674(86)90519-2. [DOI] [PubMed] [Google Scholar]
  12. Hagen G., Dennig J., Preiss A., Beato M., Suske G. Functional analyses of the transcription factor Sp4 reveal properties distinct from Sp1 and Sp3. J Biol Chem. 1995 Oct 20;270(42):24989–24994. doi: 10.1074/jbc.270.42.24989. [DOI] [PubMed] [Google Scholar]
  13. Heimberg M., Olubadewo J. O., Wilcox H. G. Plasma lipoproteins and regulation of hepatic metabolism of fatty acids in altered thyroid states. Endocr Rev. 1985 Fall;6(4):590–607. doi: 10.1210/edrv-6-4-590. [DOI] [PubMed] [Google Scholar]
  14. Jump D. B., Badin M. V., Thelen A. The CCAAT box binding factor, NF-Y, is required for thyroid hormone regulation of rat liver S14 gene transcription. J Biol Chem. 1997 Oct 31;272(44):27778–27786. doi: 10.1074/jbc.272.44.27778. [DOI] [PubMed] [Google Scholar]
  15. Krylov D., Olive M., Vinson C. Extending dimerization interfaces: the bZIP basic region can form a coiled coil. EMBO J. 1995 Nov 1;14(21):5329–5337. doi: 10.1002/j.1460-2075.1995.tb00217.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Lee Y. H., Sauer B., Johnson P. F., Gonzalez F. J. Disruption of the c/ebp alpha gene in adult mouse liver. Mol Cell Biol. 1997 Oct;17(10):6014–6022. doi: 10.1128/mcb.17.10.6014. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Lee Y. H., Williams S. C., Baer M., Sterneck E., Gonzalez F. J., Johnson P. F. The ability of C/EBP beta but not C/EBP alpha to synergize with an Sp1 protein is specified by the leucine zipper and activation domain. Mol Cell Biol. 1997 Apr;17(4):2038–2047. doi: 10.1128/mcb.17.4.2038. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Leone T. C., Cresci S., Carter M. E., Zhang Z., Lala D. S., Strauss A. W., Kelly D. P. The human medium chain Acyl-CoA dehydrogenase gene promoter consists of a complex arrangement of nuclear receptor response elements and Sp1 binding sites. J Biol Chem. 1995 Jul 7;270(27):16308–16314. doi: 10.1074/jbc.270.27.16308. [DOI] [PubMed] [Google Scholar]
  19. Levan G., Szpirer J., Szpirer C., Klinga K., Hanson C., Islam M. Q. The gene map of the Norway rat (Rattus norvegicus) and comparative mapping with mouse and man. Genomics. 1991 Jul;10(3):699–718. doi: 10.1016/0888-7543(91)90455-n. [DOI] [PubMed] [Google Scholar]
  20. Liu Y. W., Arakawa T., Yamamoto S., Chang W. C. Transcriptional activation of human 12-lipoxygenase gene promoter is mediated through Sp1 consensus sites in A431 cells. Biochem J. 1997 May 15;324(Pt 1):133–140. doi: 10.1042/bj3240133. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Majello B., De Luca P., Lania L. Sp3 is a bifunctional transcription regulator with modular independent activation and repression domains. J Biol Chem. 1997 Feb 14;272(7):4021–4026. doi: 10.1074/jbc.272.7.4021. [DOI] [PubMed] [Google Scholar]
  22. McGarry J. D., Brown N. F. The mitochondrial carnitine palmitoyltransferase system. From concept to molecular analysis. Eur J Biochem. 1997 Feb 15;244(1):1–14. doi: 10.1111/j.1432-1033.1997.00001.x. [DOI] [PubMed] [Google Scholar]
  23. Mynatt R. L., Park E. A., Thorngate F. E., Das H. K., Cook G. A. Changes in carnitine palmitoyltransferase-I mRNA abundance produced by hyperthyroidism and hypothyroidism parallel changes in activity. Biochem Biophys Res Commun. 1994 Jun 15;201(2):932–937. doi: 10.1006/bbrc.1994.1791. [DOI] [PubMed] [Google Scholar]
  24. Netzker R., Weigert C., Brand K. Role of the stimulatory proteins Sp1 and Sp3 in the regulation of transcription of the rat pyruvate kinase M gene. Eur J Biochem. 1997 Apr 1;245(1):174–181. doi: 10.1111/j.1432-1033.1997.00174.x. [DOI] [PubMed] [Google Scholar]
  25. Park E. A., Gurney A. L., Nizielski S. E., Hakimi P., Cao Z., Moorman A., Hanson R. W. Relative roles of CCAAT/enhancer-binding protein beta and cAMP regulatory element-binding protein in controlling transcription of the gene for phosphoenolpyruvate carboxykinase (GTP). J Biol Chem. 1993 Jan 5;268(1):613–619. [PubMed] [Google Scholar]
  26. Park E. A., Jerden D. C., Bahouth S. W. Regulation of phosphoenolpyruvate carboxykinase gene transcription by thyroid hormone involves two distinct binding sites in the promoter. Biochem J. 1995 Aug 1;309(Pt 3):913–919. doi: 10.1042/bj3090913. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Park E. A., Mynatt R. L., Cook G. A., Kashfi K. Insulin regulates enzyme activity, malonyl-CoA sensitivity and mRNA abundance of hepatic carnitine palmitoyltransferase-I. Biochem J. 1995 Sep 15;310(Pt 3):853–858. doi: 10.1042/bj3100853. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Park E. A., Song S., Olive M., Roesler W. J. CCAAT-enhancer-binding protein alpha (C/EBP alpha) is required for the thyroid hormone but not the retinoic acid induction of phosphoenolpyruvate carboxykinase (PEPCK) gene transcription. Biochem J. 1997 Feb 15;322(Pt 1):343–349. doi: 10.1042/bj3220343. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Park E. A., Steffen M. L., Song S., Park V. M., Cook G. A. Cloning and characterization of the promoter for the liver isoform of the rat carnitine palmitoyltransferase I (L-CPT I) gene. Biochem J. 1998 Feb 15;330(Pt 1):217–224. doi: 10.1042/bj3300217. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Roder K. H., Wolf S. S., Schweizer M. Interaction of Sp1 and NF-Y in the diet-induced regulation of the rat fatty acid synthase (FAS) gene. Biochem Soc Trans. 1997 Feb;25(1):72S–72S. doi: 10.1042/bst025072s. [DOI] [PubMed] [Google Scholar]
  31. Smale S. T. Transcription initiation from TATA-less promoters within eukaryotic protein-coding genes. Biochim Biophys Acta. 1997 Mar 20;1351(1-2):73–88. doi: 10.1016/s0167-4781(96)00206-0. [DOI] [PubMed] [Google Scholar]
  32. Tansey W. P., Schaufele F., Heslewood M., Handford C., Reudelhuber T. L., Catanzaro D. F. Distance-dependent interactions between basal, cyclic AMP, and thyroid hormone response elements in the rat growth hormone promoter. J Biol Chem. 1993 Jul 15;268(20):14906–14911. [PubMed] [Google Scholar]
  33. Thumelin S., Esser V., Charvy D., Kolodziej M., Zammit V. A., McGarry D., Girard J., Pegorier J. P. Expression of liver carnitine palmitoyltransferase I and II genes during development in the rat. Biochem J. 1994 Jun 1;300(Pt 2):583–587. doi: 10.1042/bj3000583. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Voz M. L., Peers B., Wiedig M. J., Jacquemin P., Belayew A., Martial J. A. Transcriptional regulation by triiodothyronine requires synergistic action of the thyroid receptor with another trans-acting factor. Mol Cell Biol. 1992 Sep;12(9):3991–3997. doi: 10.1128/mcb.12.9.3991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Wang D., Harrison W., Buja L. M., Elder F. F., McMillin J. B. Genomic DNA sequence, promoter expression, and chromosomal mapping of rat muscle carnitine palmitoyltransferase I. Genomics. 1998 Mar 15;48(3):314–323. doi: 10.1006/geno.1997.5184. [DOI] [PubMed] [Google Scholar]
  36. Wang N. D., Finegold M. J., Bradley A., Ou C. N., Abdelsayed S. V., Wilde M. D., Taylor L. R., Wilson D. R., Darlington G. J. Impaired energy homeostasis in C/EBP alpha knockout mice. Science. 1995 Aug 25;269(5227):1108–1112. doi: 10.1126/science.7652557. [DOI] [PubMed] [Google Scholar]
  37. Weis L., Reinberg D. Accurate positioning of RNA polymerase II on a natural TATA-less promoter is independent of TATA-binding-protein-associated factors and initiator-binding proteins. Mol Cell Biol. 1997 Jun;17(6):2973–2984. doi: 10.1128/mcb.17.6.2973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Wright K. L., Vilen B. J., Itoh-Lindstrom Y., Moore T. L., Li G., Criscitiello M., Cogswell P., Clarke J. B., Ting J. P. CCAAT box binding protein NF-Y facilitates in vivo recruitment of upstream DNA binding transcription factors. EMBO J. 1994 Sep 1;13(17):4042–4053. doi: 10.1002/j.1460-2075.1994.tb06721.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Yamazaki N., Yamanaka Y., Hashimoto Y., Shinohara Y., Shima A., Terada H. Structural features of the gene encoding human muscle type carnitine palmitoyltransferase I. FEBS Lett. 1997 Jun 16;409(3):401–406. doi: 10.1016/s0014-5793(97)00561-9. [DOI] [PubMed] [Google Scholar]
  40. van der Leij F. R., Takens J., van der Veen A. Y., Terpstra P., Kuipers J. R. Localization and intron usage analysis of the human CPT1B gene for muscle type carnitine palmitoyltransferase I. Biochim Biophys Acta. 1997 May 30;1352(2):123–128. doi: 10.1016/s0167-4781(97)00037-7. [DOI] [PubMed] [Google Scholar]

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