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. 1982;1(10):1287–1293. doi: 10.1002/j.1460-2075.1982.tb00026.x

Isolation and characterization of the rat tryptophan oxygenase gene.

W Schmid, G Scherer, U Danesch, H Zentgraf, P Matthias, C M Strange, W Röwekamp, G Schütz
PMCID: PMC553202  PMID: 6327261

Abstract

Tryptophan oxygenase (TO, EC 1.13.1.12) from rat liver is subject to glucocorticoid and developmental control. To study the mechanism of regulation, TO mRNA sequences and the chromosomal TO gene were cloned. From a cDNA library prepared from rat liver poly(A)+ RNA enriched for TO mRNA, a recombinant plasmid containing TO cDNA sequences was identified by translation of hybrid-selected RNA and immunoprecipitation with antibodies directed against TO. This cDNA clone hybridizes to a mRNA 2000 bases long that is inducible by dexamethasone. With this clone as probe we isolated from a bacteriophage lambda rat DNA library genomic clones which together span a region of 32 kilobase pairs (kb). Heteroduplex analysis revealed that the gene extends over 19 kb and is interrupted by at least 11 introns. To characterize the presumptive control region the DNA sequence around the 5' end of the TO gene was determined. S1 nuclease protection experiments revealed two separate start sites for TO mRNA transcription within this region.

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Selected References

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

  1. Benoist C., O'Hare K., Breathnach R., Chambon P. The ovalbumin gene-sequence of putative control regions. Nucleic Acids Res. 1980 Jan 11;8(1):127–142. doi: 10.1093/nar/8.1.127. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Benton W. D., Davis R. W. Screening lambdagt recombinant clones by hybridization to single plaques in situ. Science. 1977 Apr 8;196(4286):180–182. doi: 10.1126/science.322279. [DOI] [PubMed] [Google Scholar]
  3. Berk A. J., Sharp P. A. Sizing and mapping of early adenovirus mRNAs by gel electrophoresis of S1 endonuclease-digested hybrids. Cell. 1977 Nov;12(3):721–732. doi: 10.1016/0092-8674(77)90272-0. [DOI] [PubMed] [Google Scholar]
  4. Blattner F. R., Blechl A. E., Denniston-Thompson K., Faber H. E., Richards J. E., Slightom J. L., Tucker P. W., Smithies O. Cloning human fetal gamma globin and mouse alpha-type globin DNA: preparation and screening of shotgun collections. Science. 1978 Dec 22;202(4374):1279–1284. doi: 10.1126/science.725603. [DOI] [PubMed] [Google Scholar]
  5. Bonner W. M., Laskey R. A. A film detection method for tritium-labelled proteins and nucleic acids in polyacrylamide gels. Eur J Biochem. 1974 Jul 1;46(1):83–88. doi: 10.1111/j.1432-1033.1974.tb03599.x. [DOI] [PubMed] [Google Scholar]
  6. 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]
  7. Bélanger L., Frain M., Baril P., Gingras M. C., Bartkowiak J., Sala-Trepat J. M. Glucocorticosteroid suppression of alpha1-fetoprotein synthesis in developing rat liver. Evidence for selective gene repression at the transcriptional level. Biochemistry. 1981 Nov 10;20(23):6665–6672. doi: 10.1021/bi00526a022. [DOI] [PubMed] [Google Scholar]
  8. Cochet M., Chang A. C., Cohen S. N. Characterization of the structural gene and putative 5'-regulatory sequences for human proopiomelanocortin. Nature. 1982 May 27;297(5864):335–339. doi: 10.1038/297335a0. [DOI] [PubMed] [Google Scholar]
  9. Dalla-Favera R., Gelmann E. P., Gallo R. C., Wong-Staal F. A human onc gene homologous to the transforming gene (v-sis) of simian sarcoma virus. Nature. 1981 Jul 2;292(5818):31–35. doi: 10.1038/292031a0. [DOI] [PubMed] [Google Scholar]
  10. Dobner P. R., Kawasaki E. S., Yu L. Y., Bancroft F. C. Thyroid or glucocorticoid hormone induces pre-growth-hormone mRNA and its probable nuclear precursor in rat pituitary cells. Proc Natl Acad Sci U S A. 1981 Apr;78(4):2230–2234. doi: 10.1073/pnas.78.4.2230. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Donehower L. A., Huang A. L., Hager G. L. Regulatory and coding potential of the mouse mammary tumor virus long terminal redundancy. J Virol. 1981 Jan;37(1):226–238. doi: 10.1128/jvi.37.1.226-238.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Fasel N., Pearson K., Buetti E., Diggelmann H. The region of mouse mammary tumor virus DNA containing the long terminal repeat includes a long coding sequence and signals for hormonally regulated transcription. EMBO J. 1982;1(1):3–7. doi: 10.1002/j.1460-2075.1982.tb01115.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Franz J. M., Knox W. E. The effect of development and hydrocortisone on tryptophan oxygenase, formamidase, and tyrosine aminotransferase in the livers of young rats. Biochemistry. 1967 Nov;6(11):3464–3471. doi: 10.1021/bi00863a017. [DOI] [PubMed] [Google Scholar]
  14. Govindan M. V., Spiess E., Majors J. Purified glucocorticoid receptor-hormone complex from rat liver cytosol binds specifically to cloned mouse mammary tumor virus long terminal repeats in vitro. Proc Natl Acad Sci U S A. 1982 Sep;79(17):5157–5161. doi: 10.1073/pnas.79.17.5157. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Grez M., Land H., Giesecke K., Schütz G., Jung A., Sippel A. E. Multiple mRNAs are generated from the chicken lysozyme gene. Cell. 1981 Sep;25(3):743–752. doi: 10.1016/0092-8674(81)90182-3. [DOI] [PubMed] [Google Scholar]
  16. Hagenbüchle O., Tosi M., Schibler U., Bovey R., Wellauer P. K., Young R. A. Mouse liver and salivary gland alpha-amylase mRNAs differ only in 5' non-translated sequences. Nature. 1981 Feb 19;289(5799):643–646. doi: 10.1038/289643a0. [DOI] [PubMed] [Google Scholar]
  17. Herrmann R., Neugebauer K., Zentgraf H., Schaller H. Transposition of a DNA sequence determining kanamycin resistance into the single-stranded genome of bacteriophage fd. Mol Gen Genet. 1978 Feb 16;159(2):171–178. doi: 10.1007/BF00270890. [DOI] [PubMed] [Google Scholar]
  18. Hofer E., Land H., Sekeris C. E., Morris H. P. Messenger RNA activities for two liver enzymes, tyrosine aminotransferase and tryptophan oxygenase, in Morris hepatoma 5123C and 9618A and in HTC cells. Correlation with enzyme activities. Eur J Biochem. 1978 Nov 2;91(1):223–229. doi: 10.1111/j.1432-1033.1978.tb20955.x. [DOI] [PubMed] [Google Scholar]
  19. Hofer E., Sekeris C. E. The messenger RNA's for the liver enzymes tyrosine aminotransferase and tryptophan oxygenase contain 40-50 % non-coding sequences. Biochem Biophys Res Commun. 1977 Jul 11;77(1):352–360. doi: 10.1016/s0006-291x(77)80204-0. [DOI] [PubMed] [Google Scholar]
  20. KENNEY F. T. Induction of tyrosine-alpha-ketoglutarate transaminase in rat liver. IV. Evidence for an increase in the rate of enzyme synthesis. J Biol Chem. 1962 Nov;237:3495–3498. [PubMed] [Google Scholar]
  21. KNOX W. E., MEHLER A. H. The adaptive increase of the tryptophan peroxidase-oxidase system of liver. Science. 1951 Mar 2;113(2931):237–238. doi: 10.1126/science.113.2931.237. [DOI] [PubMed] [Google Scholar]
  22. Kennedy N., Knedlitschek G., Groner B., Hynes N. E., Herrlich P., Michalides R., van Ooyen A. J. Long terminal repeats of endogenous mouse mammary tumour virus contain a long open reading frame which extends into adjacent sequences. Nature. 1982 Feb 18;295(5850):622–624. doi: 10.1038/295622a0. [DOI] [PubMed] [Google Scholar]
  23. Killewich L. A., Feigelson P. Developmental control of messenger RNA for hepatic tryptophan 2,3-dioxygenase. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5392–5396. doi: 10.1073/pnas.74.12.5392. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Koller B., Delius H., Bünemann H., Müller W. The isolation of DNA from agarose gels by electrophoretic elution onto malachite green-polyacrylamide columns. Gene. 1978 Nov;4(3):227–239. doi: 10.1016/0378-1119(78)90020-3. [DOI] [PubMed] [Google Scholar]
  25. LIN E. C., KNOX W. E. Specificity of the adaptive response to tyrosine-alpha-ketoglutarate transaminase in the rat. J Biol Chem. 1958 Nov;233(5):1186–1189. [PubMed] [Google Scholar]
  26. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  27. LeMeur M., Glanville N., Mandel J. L., Gerlinger P., Palmiter R., Chambon P. The ovalbumin gene family: hormonal control of X and Y gene transcription and mRNA accumulation. Cell. 1981 Feb;23(2):561–571. doi: 10.1016/0092-8674(81)90152-5. [DOI] [PubMed] [Google Scholar]
  28. Lindenmaier W., Nguyen-Huu M. C., Lurz R., Stratmann M., Blin N., Wurtz T., Hauser H. J., Sippel A. E., Schütz G. Arrangement of coding and intervening sequences of chicken lysozyme gene. Proc Natl Acad Sci U S A. 1979 Dec;76(12):6196–6200. doi: 10.1073/pnas.76.12.6196. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Maxam A. M., Gilbert W. Sequencing end-labeled DNA with base-specific chemical cleavages. Methods Enzymol. 1980;65(1):499–560. doi: 10.1016/s0076-6879(80)65059-9. [DOI] [PubMed] [Google Scholar]
  30. McMaster G. K., Carmichael G. G. Analysis of single- and double-stranded nucleic acids on polyacrylamide and agarose gels by using glyoxal and acridine orange. Proc Natl Acad Sci U S A. 1977 Nov;74(11):4835–4838. doi: 10.1073/pnas.74.11.4835. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. NEMETH A. M. Mechanisms controlling changes in tryptophan peroxidase activity in developing mammalian liver. J Biol Chem. 1959 Nov;234:2921–2924. [PubMed] [Google Scholar]
  32. Page G. S., Smith S., Goodman H. M. DNA sequence of the rat growth hormone gene: location of the 5' terminus of the growth hormone mRNA and identification of an internal transposon-like element. Nucleic Acids Res. 1981 May 11;9(9):2087–2104. doi: 10.1093/nar/9.9.2087. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Ramanarayanan-Murthy L., Colman P. D., Morris H. P., Feigelson P. Pretranslational control of tryptophan oxygenase levels in Morris hepatoma and host liver. Cancer Res. 1976 Oct;36(10):3594–3599. [PubMed] [Google Scholar]
  34. Ricciardi R. P., Miller J. S., Roberts B. E. Purification and mapping of specific mRNAs by hybridization-selection and cell-free translation. Proc Natl Acad Sci U S A. 1979 Oct;76(10):4927–4931. doi: 10.1073/pnas.76.10.4927. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Rigby P. W., Dieckmann M., Rhodes C., Berg P. Labeling deoxyribonucleic acid to high specific activity in vitro by nick translation with DNA polymerase I. J Mol Biol. 1977 Jun 15;113(1):237–251. doi: 10.1016/0022-2836(77)90052-3. [DOI] [PubMed] [Google Scholar]
  36. Roewekamp W. G., Hofer E., Sekeris C. E. Translation of mRNA from rat-liver polysomes into tyrosine aminotransferase and tryptophan oxygenase in a protein-synthesizing system from wheat germ. Effects of cortisol on the translatable levels of mRNA for these two enzymes. Eur J Biochem. 1976 Nov 1;70(1):259–268. doi: 10.1111/j.1432-1033.1976.tb10977.x. [DOI] [PubMed] [Google Scholar]
  37. Rowekamp W., Firtel R. A. Isolation of developmentally regulated genes from Dictyostelium. Dev Biol. 1980 Oct;79(2):409–418. doi: 10.1016/0012-1606(80)90126-8. [DOI] [PubMed] [Google Scholar]
  38. SCHIMKE R. T., SWEENEY E. W., BERLIN C. M. THE ROLES OF SYNTHESIS AND DEGRADATION IN THE CONTROL OF RAT LIVER TRYPTOPHAN PYRROLASE. J Biol Chem. 1965 Jan;240:322–331. [PubMed] [Google Scholar]
  39. Sargent T. D., Wu J. R., Sala-Trepat J. M., Wallace R. B., Reyes A. A., Bonner J. The rat serum albumin gene: analysis of cloned sequences. Proc Natl Acad Sci U S A. 1979 Jul;76(7):3256–3260. doi: 10.1073/pnas.76.7.3256. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Sargent T. D., Yang M., Bonner J. Nucleotide sequence of cloned rat serum albumin messenger RNA. Proc Natl Acad Sci U S A. 1981 Jan;78(1):243–246. doi: 10.1073/pnas.78.1.243. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Schutz G., Beato M., Feigelson P. Messenger RNA for hepatic tryptophan oxygenase: its partial purification, its translation in a heterologous cell-free system, and its control by glucocorticoid hormones. Proc Natl Acad Sci U S A. 1973 Apr;70(4):1218–1221. doi: 10.1073/pnas.70.4.1218. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Schutz G., Killewich L., Chen G., Feigelson P. Control of the mRNA for hepatic tryptophan oxygenase during hormonal and substrate induction. Proc Natl Acad Sci U S A. 1975 Mar;72(3):1017–1020. doi: 10.1073/pnas.72.3.1017. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. Tomizawa J. I., Ohmori H., Bird R. E. Origin of replication of colicin E1 plasmid DNA. Proc Natl Acad Sci U S A. 1977 May;74(5):1865–1869. doi: 10.1073/pnas.74.5.1865. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. Tsukada T., Watanabe Y., Nakai Y., Imura H., Nakanishi S., Numa S. Repetitive DNA sequences in the human corticotropin-beta-lipotrophin precursor gene region: Alu family members. Nucleic Acids Res. 1982 Mar 11;10(5):1471–1479. doi: 10.1093/nar/10.5.1471. [DOI] [PMC free article] [PubMed] [Google Scholar]
  45. Weaver R. F., Weissmann C. Mapping of RNA by a modification of the Berk-Sharp procedure: the 5' termini of 15 S beta-globin mRNA precursor and mature 10 s beta-globin mRNA have identical map coordinates. Nucleic Acids Res. 1979 Nov 10;7(5):1175–1193. doi: 10.1093/nar/7.5.1175. [DOI] [PMC free article] [PubMed] [Google Scholar]

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