Molecular and General Genetics MGG

, Volume 248, Issue 6, pp 755–766 | Cite as

Theraspberry locus encodesDrosophila inosine monophosphate dehydrogenase

  • Roger Slee
  • Mary Bownes
Original Paper


Investigation of an enhancer-trap line exhibiting testis-specificβ-galactosidase expression led to the isolation of theDrosophila gene encoding inosine monophosphate dehydrogenase (IMPD), the rate-limiting enzyme in guanine nucleotide synthesis, which has been implicated in cell cycle control and malignant transformation. Northern and in situ hybridization analysis demonstrated that the gene has a complex expression pattern involving several independently regulated transcripts. Two ubiquitous, but highly ovary enriched, transcripts of 2.5 and 1.9 kb are expressed in the nurse cells and delivered to the oocyte, whilst a 0.9 kb transcript is found exclusively in the testis. The 2.5 kb transcript encodes a 58 kDa protein, which is highly similar in length and sequence to mouse and human IMPDs and is presumably required for GTP synthesis during early embryogenesis. Over-expression of this cDNA inEscherichia coli yielded a product of the predicted size, which was demonstrated to possess IMPD activity in a spectrophotometric assay. The coding capacity of the other transcripts is currently uncertain. We present evidence that IMPD is the product of theraspberry (ras) locus at 9E and the functions of the gene are discussed in relation to the phenotypes ofras mutants.

Key words

Drosophila Inosine monophosphate dehydrogenase Guanine nucleotide metabolism 


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. Biggs J, Hersperger E, Steeg PS, Liotta LA, Shearn A (1990) A Drosophila gene that is homologous to a mammalian gene associated with tumor metastasis codes for a nucleoside diphosphate kinase. Cell 63:933–940Google Scholar
  2. Bourne HR, Sanders DA, McCormick F (1990) The GTPase superfamily: a conserved switch for diverse cell functions. Nature 348:125–131Google Scholar
  3. Bownes M (1990) Preferential insertion of P-elements into genes expressed in the germline ofDrosophila melanogaster. Mol Gen Genet 222:457–460Google Scholar
  4. Brand AH, Perrimon N (1993) Targeted gene expression as a means of altering cell fates and generating dominant phenotypes. Development 118:127–138Google Scholar
  5. Cavener D (1987) Comparison of the consensus sequence flanking translational start sites in Drosophila and vertebrates. Nucleic Acids Res 4:1353–1361Google Scholar
  6. Cohen MB, Sadee W (1983) Contributions of the depletions of guanine and adenine nucleotides to the toxicity of purine starvation in the mouse T-lymphoma cell-line. Cancer Res 43:1587–1591Google Scholar
  7. Collart F, Huberman E (1988) Cloning and sequence analysis of the human and chinese hamster inosine-5′-monophosphate dehydrogenase cDNAs. J Biol Chem 263:15769–15772Google Scholar
  8. Devereux J, Haeberli P, Smithies O (1984) A comprehensive set of sequence analysis programs for the VAX. Nucleic Acids Res 12:387–395Google Scholar
  9. Falk D, Nash D (1974) Sex-linked auxotrophic and putative auxotrophic mutants ofDrosophila melanogaster. Genetics 76:755–766Google Scholar
  10. Feinberg AP, Vogelstein B (1983) A technique for radiolabelling DNA restriction endonuclease fragments to high specific activity. Anal Biochem 132:6Google Scholar
  11. Frohman MA, Dush MK, Martin GR (1988) Rapid production of full length cDNAs from rare transcripts using a single genespecific oligonucleotide primer. Proc Natl Acad Sci USA 85:8998–9002Google Scholar
  12. Hupe DJ, Azzolina BA, Behrens ND (1986) IMP dehydrogenase from the intracellular parasitic protozoanEimeria tenella and its inhibition by mycophenolic acid. J Biol Chem 261:8363–8369Google Scholar
  13. Jackson RC, Weber G, Morris H (1975) IMP dehydrogenase, an enzyme linked with proliferation and malignancy. Nature 256:331–333Google Scholar
  14. Johnson MM, Woloshyn EP, Nash D (1979) Cytogenetic localisation of the purine 1 and guanosine 1 loci ofDrosophila melanogaster; the purine 1 locus specifies a vital function. Mol Gen Genet 174:287–292Google Scholar
  15. Jowett T (1986) Preparation of nucleic acids. In: Roberts DB (ed)Drosophila a practical approach. IRL Press, Oxford, pp 275–286Google Scholar
  16. Kimura N, Johnson G (1983) Increased membrane-associated, nucleoside diphosphate kinase-activity as a possible basis for enhanced guanine nucleotide-dependent adenylate-cyclase activity induced by picolinic-acid treatment of simian-virus-40-transformed normal rat-kidney cells. J Biol Chem 258:2609–2617Google Scholar
  17. Kimura N, Shimada N (1988) Membrane associated nucleoside diphosphate kinase from rat liver-purification, characterisation, and comparison with cytosolic enzyme. J Biol Chem 263:4647–4653Google Scholar
  18. King RC (1970) Ovarian development inDrosophila melanogaster. Academic Press, New YorkGoogle Scholar
  19. Kobayashi T, Simizu T (1976) Roles of nucleoside diphosphate kinases in microtubule assembly. J Biochem 79:1357–1364Google Scholar
  20. Konno Y, Natsumeda Y, Nagai M, Yamaji Y, Ohno S, Suzuki K, Weber G (1991) Expression of human IMP dehydrogenases types I and II inEscherichia coli and distribution in human normal lymphocytes and leukaemia cell lines. J Biol Chem 266:506–509Google Scholar
  21. Lindsley DL, Zimm GG (1992) The genome ofDrosophila melanogaster. Academic Press, New YorkGoogle Scholar
  22. Nash D, Woloshyn EP, Mehl YM, Janca FC (1981) Pleiotropic, recessive-lethal mutants associated with purine metabolism in Drosophila melanogaster. Can J Genet Cytol:23:411–423Google Scholar
  23. Nash D, Hu S, Leonard NJ, Tiong SYK, Fillips D (1994) Theraspberry locus ofDrosophila melanogaster includes an inosine monophosphate dehydrogenase like coding sequence. Genome 37:333–344Google Scholar
  24. Natsumeda Y, Ikegami T, Murayama K, Weber G (1988) De novo guanylate synthesis in the commitment to replication in hepatoma 3924A cells. Cancer Res 48:507–511Google Scholar
  25. Natsumeda Y, Ohno S, Kawasaki H, Konno Y, Weber G, Suzuki K (1990) Two distinct cDNAs for human IMP dehydrogenase. J. Biochem 265:5292–5295Google Scholar
  26. Ohtsuki K, Yokoyama M (1987) Direct activation of guanine-nucleotide binding-proteins through high energy phosphate transfer by nucleoside diphosphate kinase. Biochem Biophys Res Commun 148:300–307Google Scholar
  27. Proffitt RT, Pathak VK, Villacorte DG, Presant CA (1983) Sensitive radiochemical assay for inosine 5′-monophosphate deydrogenase and determination of activity in murine tumor and tissue extracts. Cancer Res 43:1620–1623Google Scholar
  28. Sambrook J, Fritsch E, Maniatis T (1989) Molecular cloning: a laboratory manual. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New YorkGoogle Scholar
  29. Satoh T, Nakafuku M, Kaziro Y (1992) Function of Ras as a molecular switch in signal transduction. J Biol Chem 267:24149–24152Google Scholar
  30. Sherley JL (1991) Guanine nucleotide biosynthesis is regulated by the cellular p53 concentration. J Biochem 266:24815–24828Google Scholar
  31. Shimura K, Okada M, Shiraki H, Nakagawa H (1983) IMP dehydrogenase 1. Studies on regulatory properties of crude tissueextracts based on an improved assay method. J Biochem 94:1595–1603Google Scholar
  32. Slee R (1992) Cloning and characterisation ofDrosophila IMP dehydrogenase. PhD Thesis, Institute of Cell and Molecular Biology, University of EdinburghGoogle Scholar
  33. Steeg PS, Bevilacqua G, Kopper L, Thorgeirsson U, Talmadge J, Liotta L, Sobel M (1988) Evidence for a novel gene associated with low tumor metastatic potential. J Natl Cancer Inst 80:200–204Google Scholar
  34. Tautz D, Pfeifle C (1989) A non-radioactive in situ hybridisation method for the localisation of specific RNAs in Drosophila embryos reveals translational control of the segmentation genehunchback. Chromosoma 98:81–85Google Scholar
  35. Tiedeman A, Smith J (1985) Nucleotide sequence of the guaB locus encoding IMP dehydrogenase ofEscherichia coli K12. Nucleic Acids Res 13:1303–1316Google Scholar
  36. Tiedeman A, Smith J (1991) Isolation and sequence of a cDNA encoding mouse IMP dehydrogenase. Gene 97:289–293Google Scholar
  37. Verham R, Meek T, Hedstrom L, Wang C (1987) Purification, characterisation and kinetic analysis of inosine 5′-monophosphate dehydrogenase ofTritrichonomas foetus. Mol Biochem Parasitol 24:1–12Google Scholar
  38. Weber G (1983) Biochemical strategy of cancer cells and the design of chemotherapy. Cancer Res 43:3466–3492Google Scholar
  39. Wilson K, Collart F, Huberman E, Stringer J, Ullman B (1991) Amplification and molecular cloning of the IMP dehydrogenase gene ofLeishmania donovani. J Biol Chem 266:1665–1671Google Scholar
  40. Woods DF, Bryant PJ (1991) The discs-large tumour suppressor gene of Drosophila encodes a guanylate kinase homolog localised at septate junctions. Cell 66:451–464Google Scholar

Copyright information

© Springer-Verlag 1995

Authors and Affiliations

  • Roger Slee
    • 1
  • Mary Bownes
    • 1
  1. 1.Institute of Cell and Molecular BiologyUniversity of EdinburghEdinburghUK

Personalised recommendations