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. 2003 Mar 15;370(Pt 3):771–784. doi: 10.1042/BJ20021445

Genomic organization and transcriptional analysis of the human l-glutaminase gene.

Cristina Pérez-Gómez 1, José M Matés 1, Pedro M Gómez-Fabre 1, Antonio del Castillo-Olivares 1, Francisco J Alonso 1, Javier Márquez 1
PMCID: PMC1223212  PMID: 12444921

Abstract

In mammals, glutaminase (GA) is expressed in most tissues, but the regulation of organ-specific expression is largely unknown. Therefore, as an essential step towards studying the regulation of GA expression, the human liver-type GA (hLGA) gene has been characterized. LGA genomic sequences were isolated using the genome walking technique. Analysis and comparison of these sequences with two LGA cDNA clones and the Human Genome Project database, allowed the determination of the genomic organization of the LGA gene. The gene has 18 exons and is approx. 18 kb long. All exon/intron junction sequences conform to the GT/AG rule. Progressive deletion analysis of LGA promoter-luciferase constructs indicated that the core promoter is located between nt -141 and +410, with several potential regulatory elements: CAAT, GC, TATA-like, Ras-responsive element binding protein and specificity protein 1 (Sp1) sites. The minimal promoter was mapped within +107 and +410, where only an Sp1 binding site is present. Mutation experiments suggested that two CAAT recognition elements near the transcription-initiation site (-138 and -87), play a crucial role for optimal promoter activity. Electrophoretic mobility-shift assays confirmed the importance of CAAT- and TATA-like boxes to enhance basal transcription, and demonstrated that HNF-1 motif is a significant distal element for transcriptional regulation of the hLGA gene.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Aledo J. C., Gómez-Fabre P. M., Olalla L., Márquez J. Identification of two human glutaminase loci and tissue-specific expression of the two related genes. Mamm Genome. 2000 Dec;11(12):1107–1110. doi: 10.1007/s003350010190. [DOI] [PubMed] [Google Scholar]
  2. Aledo J. C., Segura J. A., Medina M. A., Alonso F. J., Núez de Castro I., Márquez J. Phosphate-activated glutaminase expression during tumor development. FEBS Lett. 1994 Mar 14;341(1):39–42. doi: 10.1016/0014-5793(94)80236-x. [DOI] [PubMed] [Google Scholar]
  3. Bode B. P., Souba W. W. Modulation of cellular proliferation alters glutamine transport and metabolism in human hepatoma cells. Ann Surg. 1994 Oct;220(4):411–424. doi: 10.1097/00000658-199410000-00001. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Boisclair Y. R., Brown A. L., Casola S., Rechler M. M. Three clustered Sp1 sites are required for efficient transcription of the TATA-less promoter of the gene for insulin-like growth factor-binding protein-2 from the rat. J Biol Chem. 1993 Nov 25;268(33):24892–24901. [PubMed] [Google Scholar]
  5. Breathnach R., Chambon P. Organization and expression of eucaryotic split genes coding for proteins. Annu Rev Biochem. 1981;50:349–383. doi: 10.1146/annurev.bi.50.070181.002025. [DOI] [PubMed] [Google Scholar]
  6. Chung-Bok M. I., Vincent N., Jhala U., Watford M. Rat hepatic glutaminase: identification of the full coding sequence and characterization of a functional promoter. Biochem J. 1997 May 15;324(Pt 1):193–200. doi: 10.1042/bj3240193. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Colquhoun A., Newsholme E. A. Aspects of glutamine metabolism in human tumour cells. Biochem Mol Biol Int. 1997 Mar;41(3):583–596. doi: 10.1080/15216549700201621. [DOI] [PubMed] [Google Scholar]
  8. Curthoys N. P., Watford M. Regulation of glutaminase activity and glutamine metabolism. Annu Rev Nutr. 1995;15:133–159. doi: 10.1146/annurev.nu.15.070195.001025. [DOI] [PubMed] [Google Scholar]
  9. Darlington G. J., Ross S. E., MacDougald O. A. The role of C/EBP genes in adipocyte differentiation. J Biol Chem. 1998 Nov 13;273(46):30057–30060. doi: 10.1074/jbc.273.46.30057. [DOI] [PubMed] [Google Scholar]
  10. Elgadi K. M., Meguid R. A., Qian M., Souba W. W., Abcouwer S. F. Cloning and analysis of unique human glutaminase isoforms generated by tissue-specific alternative splicing. Physiol Genomics. 1999 Aug 31;1(2):51–62. doi: 10.1152/physiolgenomics.1999.1.2.51. [DOI] [PubMed] [Google Scholar]
  11. Friedman A. D., Landschulz W. H., McKnight S. L. CCAAT/enhancer binding protein activates the promoter of the serum albumin gene in cultured hepatoma cells. Genes Dev. 1989 Sep;3(9):1314–1322. doi: 10.1101/gad.3.9.1314. [DOI] [PubMed] [Google Scholar]
  12. Gómez-Fabre P. M., Aledo J. C., Del Castillo-Olivares A., Alonso F. J., Núez De Castro I., Campos J. A., Márquez J. Molecular cloning, sequencing and expression studies of the human breast cancer cell glutaminase. Biochem J. 2000 Jan 15;345(Pt 2):365–375. [PMC free article] [PubMed] [Google Scholar]
  13. Kovacevic Z., McGivan J. D. Mitochondrial metabolism of glutamine and glutamate and its physiological significance. Physiol Rev. 1983 Apr;63(2):547–605. doi: 10.1152/physrev.1983.63.2.547. [DOI] [PubMed] [Google Scholar]
  14. Krebs H. A. Metabolism of amino-acids: The synthesis of glutamine from glutamic acid and ammonia, and the enzymic hydrolysis of glutamine in animal tissues. Biochem J. 1935 Aug;29(8):1951–1969. doi: 10.1042/bj0291951. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Laterza O. F., Hansen W. R., Taylor L., Curthoys N. P. Identification of an mRNA-binding protein and the specific elements that may mediate the pH-responsive induction of renal glutaminase mRNA. J Biol Chem. 1997 Sep 5;272(36):22481–22488. doi: 10.1074/jbc.272.36.22481. [DOI] [PubMed] [Google Scholar]
  16. Lobo C., Ruiz-Bellido M. A., Aledo J. C., Márquez J., Núez De Castro I., Alonso F. J. Inhibition of glutaminase expression by antisense mRNA decreases growth and tumourigenicity of tumour cells. Biochem J. 2000 Jun 1;348(Pt 2):257–261. [PMC free article] [PubMed] [Google Scholar]
  17. Lohmann R., Souba W. W., Bode B. P. Rat liver endothelial cell glutamine transporter and glutaminase expression contrast with parenchymal cells. Am J Physiol. 1999 Mar;276(3 Pt 1):G743–G750. doi: 10.1152/ajpgi.1999.276.3.G743. [DOI] [PubMed] [Google Scholar]
  18. Makałowski W., Mitchell G. A., Labuda D. Alu sequences in the coding regions of mRNA: a source of protein variability. Trends Genet. 1994 Jun;10(6):188–193. doi: 10.1016/0168-9525(94)90254-2. [DOI] [PubMed] [Google Scholar]
  19. Marston Steven B., Ingwall Joanne S., Glueck Susan B. Calcium, contractions, and tropomyosin Focus on "divergent abnormal muscle relaxation by hypertrophic cardiomyopathy and nemaline myopathy mutant tropomyosins". Physiol Genomics. 2002;9(2):57–58. doi: 10.1152/physiolgenomics.00038.2002. [DOI] [PubMed] [Google Scholar]
  20. Matsuno T., Goto I. Glutaminase and glutamine synthetase activities in human cirrhotic liver and hepatocellular carcinoma. Cancer Res. 1992 Mar 1;52(5):1192–1194. [PubMed] [Google Scholar]
  21. Matés José M., Pérez-Gómez Cristina, Núez de Castro Ignacio, Asenjo Maite, Márquez Javier. Glutamine and its relationship with intracellular redox status, oxidative stress and cell proliferation/death. Int J Biochem Cell Biol. 2002 May;34(5):439–458. doi: 10.1016/s1357-2725(01)00143-1. [DOI] [PubMed] [Google Scholar]
  22. Medina M. A., Sánchez-Jiménez F., Márquez J., Rodríguez Quesada A., Núez de Castro I. Relevance of glutamine metabolism to tumor cell growth. Mol Cell Biochem. 1992 Jul 6;113(1):1–15. doi: 10.1007/BF00230880. [DOI] [PubMed] [Google Scholar]
  23. Nagase T., Ishikawa K., Suyama M., Kikuno R., Hirosawa M., Miyajima N., Tanaka A., Kotani H., Nomura N., Ohara O. Prediction of the coding sequences of unidentified human genes. XII. The complete sequences of 100 new cDNA clones from brain which code for large proteins in vitro. DNA Res. 1998 Dec 31;5(6):355–364. doi: 10.1093/dnares/5.6.355. [DOI] [PubMed] [Google Scholar]
  24. Norris J., Fan D., Aleman C., Marks J. R., Futreal P. A., Wiseman R. W., Iglehart J. D., Deininger P. L., McDonnell D. P. Identification of a new subclass of Alu DNA repeats which can function as estrogen receptor-dependent transcriptional enhancers. J Biol Chem. 1995 Sep 29;270(39):22777–22782. doi: 10.1074/jbc.270.39.22777. [DOI] [PubMed] [Google Scholar]
  25. Olalla L., Aledo J. C., Bannenberg G., Márquez J. The C-terminus of human glutaminase L mediates association with PDZ domain-containing proteins. FEBS Lett. 2001 Jan 19;488(3):116–122. doi: 10.1016/s0014-5793(00)02373-5. [DOI] [PubMed] [Google Scholar]
  26. Olalla Lucía, Gutiérrez Antonia, Campos José A., Khan Zafar U., Alonso Francisco J., Segura Juan A., Márquez Javier, Aledo J. Carlos. Nuclear localization of L-type glutaminase in mammalian brain. J Biol Chem. 2002 Aug 5;277(41):38939–38944. doi: 10.1074/jbc.C200373200. [DOI] [PubMed] [Google Scholar]
  27. 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]
  28. Quandt K., Frech K., Karas H., Wingender E., Werner T. MatInd and MatInspector: new fast and versatile tools for detection of consensus matches in nucleotide sequence data. Nucleic Acids Res. 1995 Dec 11;23(23):4878–4884. doi: 10.1093/nar/23.23.4878. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Sanger F., Nicklen S., Coulson A. R. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. doi: 10.1073/pnas.74.12.5463. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Shapiro R. A., Farrell L., Srinivasan M., Curthoys N. P. Isolation, characterization, and in vitro expression of a cDNA that encodes the kidney isoenzyme of the mitochondrial glutaminase. J Biol Chem. 1991 Oct 5;266(28):18792–18796. [PubMed] [Google Scholar]
  31. Shen M. R., Batzer M. A., Deininger P. L. Evolution of the master Alu gene(s). J Mol Evol. 1991 Oct;33(4):311–320. doi: 10.1007/BF02102862. [DOI] [PubMed] [Google Scholar]
  32. Smith E. M., Watford M. Molecular cloning of a cDNA for rat hepatic glutaminase. Sequence similarity to kidney-type glutaminase. J Biol Chem. 1990 Jun 25;265(18):10631–10636. [PubMed] [Google Scholar]
  33. Taylor L., Liu X., Newsome W., Shapiro R. A., Srinivasan M., Curthoys N. P. Isolation and characterization of the promoter region of the rat kidney-type glutaminase gene. Biochim Biophys Acta. 2001 Mar 19;1518(1-2):132–136. doi: 10.1016/s0167-4781(01)00183-x. [DOI] [PubMed] [Google Scholar]

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