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Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1988 Oct;85(19):7134–7138. doi: 10.1073/pnas.85.19.7134

Cloning of a Drosophila melanogaster guanine nucleotide regulatory protein beta-subunit gene and characterization of its expression during development.

S Yarfitz 1, N M Provost 1, J B Hurley 1
PMCID: PMC282138  PMID: 3140235

Abstract

A Drosophila melanogaster gene encoding a protein with greater than 80% sequence identity to the beta subunits of mammalian guanine nucleotide-binding regulatory proteins (G proteins) has been cloned. The gene, which was mapped to 13F on the X chromosome by in situ hybridization, was cloned from a Drosophila genomic library by using a bovine transducin beta-subunit cDNA probe. Genomic DNA blot hybridization analysis indicated that there is a single Drosophila G-protein beta-subunit gene. Multiple transcripts were detected throughout development; in adult flies the mRNA is expressed at higher levels in heads than in bodies. The proposed coding region is uninterrupted by introns, but there is evidence for differential mRNA splicing in the 5' nontranslated region.

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Selected References

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  1. Baer K. M., Saibil H. R. Light- and GTP-activated hydrolysis of phosphatidylinositol bisphosphate in squid photoreceptor membranes. J Biol Chem. 1988 Jan 5;263(1):17–20. [PubMed] [Google Scholar]
  2. Blumenfeld A., Erusalimsky J., Heichal O., Selinger Z., Minke B. Light-activated guanosinetriphosphatase in Musca eye membranes resembles the prolonged depolarizing afterpotential in photoreceptor cells. Proc Natl Acad Sci U S A. 1985 Oct;82(20):7116–7120. doi: 10.1073/pnas.82.20.7116. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Brown J. E., Rubin L. J., Ghalayini A. J., Tarver A. P., Irvine R. F., Berridge M. J., Anderson R. E. myo-Inositol polyphosphate may be a messenger for visual excitation in Limulus photoreceptors. Nature. 1984 Sep 13;311(5982):160–163. doi: 10.1038/311160a0. [DOI] [PubMed] [Google Scholar]
  4. Cathala G., Savouret J. F., Mendez B., West B. L., Karin M., Martial J. A., Baxter J. D. A method for isolation of intact, translationally active ribonucleic acid. DNA. 1983;2(4):329–335. doi: 10.1089/dna.1983.2.329. [DOI] [PubMed] [Google Scholar]
  5. Cavener D. R. Comparison of the consensus sequence flanking translational start sites in Drosophila and vertebrates. Nucleic Acids Res. 1987 Feb 25;15(4):1353–1361. doi: 10.1093/nar/15.4.1353. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Codina J., Stengel D., Woo S. L., Birnbaumer L. Beta-subunits of the human liver Gs/Gi signal-transducing proteins and those of bovine retinal rod cell transducin are identical. FEBS Lett. 1986 Oct 27;207(2):187–192. doi: 10.1016/0014-5793(86)81486-7. [DOI] [PubMed] [Google Scholar]
  7. Devary O., Heichal O., Blumenfeld A., Cassel D., Suss E., Barash S., Rubinstein C. T., Minke B., Selinger Z. Coupling of photoexcited rhodopsin to inositol phospholipid hydrolysis in fly photoreceptors. Proc Natl Acad Sci U S A. 1987 Oct;84(19):6939–6943. doi: 10.1073/pnas.84.19.6939. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Dietzel C., Kurjan J. The yeast SCG1 gene: a G alpha-like protein implicated in the a- and alpha-factor response pathway. Cell. 1987 Sep 25;50(7):1001–1010. doi: 10.1016/0092-8674(87)90166-8. [DOI] [PubMed] [Google Scholar]
  9. Evans T., Fawzi A., Fraser E. D., Brown M. L., Northup J. K. Purification of a beta 35 form of the beta gamma complex common to G-proteins from human placental membranes. J Biol Chem. 1987 Jan 5;262(1):176–181. [PubMed] [Google Scholar]
  10. Fong H. K., Amatruda T. T., 3rd, Birren B. W., Simon M. I. Distinct forms of the beta subunit of GTP-binding regulatory proteins identified by molecular cloning. Proc Natl Acad Sci U S A. 1987 Jun;84(11):3792–3796. doi: 10.1073/pnas.84.11.3792. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Fong H. K., Hurley J. B., Hopkins R. S., Miake-Lye R., Johnson M. S., Doolittle R. F., Simon M. I. Repetitive segmental structure of the transducin beta subunit: homology with the CDC4 gene and identification of related mRNAs. Proc Natl Acad Sci U S A. 1986 Apr;83(7):2162–2166. doi: 10.1073/pnas.83.7.2162. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Fung B. K. Characterization of transducin from bovine retinal rod outer segments. I. Separation and reconstitution of the subunits. J Biol Chem. 1983 Sep 10;258(17):10495–10502. [PubMed] [Google Scholar]
  13. Ganetzky B., Wu C. F. Neurogenetics of membrane excitability in Drosophila. Annu Rev Genet. 1986;20:13–44. doi: 10.1146/annurev.ge.20.120186.000305. [DOI] [PubMed] [Google Scholar]
  14. Gao B., Gilman A. G., Robishaw J. D. A second form of the beta subunit of signal-transducing G proteins. Proc Natl Acad Sci U S A. 1987 Sep;84(17):6122–6125. doi: 10.1073/pnas.84.17.6122. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Gao B., Mumby S., Gilman A. G. The G protein beta 2 complementary DNA encodes the beta 35 subunit. J Biol Chem. 1987 Dec 25;262(36):17254–17257. [PubMed] [Google Scholar]
  16. Geisow M. J. Common domain structure of Ca2+ and lipid-binding proteins. FEBS Lett. 1986 Jul 14;203(1):99–103. doi: 10.1016/0014-5793(86)81445-4. [DOI] [PubMed] [Google Scholar]
  17. Gilman A. G. G proteins: transducers of receptor-generated signals. Annu Rev Biochem. 1987;56:615–649. doi: 10.1146/annurev.bi.56.070187.003151. [DOI] [PubMed] [Google Scholar]
  18. Hattori M., Sakaki Y. Dideoxy sequencing method using denatured plasmid templates. Anal Biochem. 1986 Feb 1;152(2):232–238. doi: 10.1016/0003-2697(86)90403-3. [DOI] [PubMed] [Google Scholar]
  19. Hurley J. B., Fong H. K., Teplow D. B., Dreyer W. J., Simon M. I. Isolation and characterization of a cDNA clone for the gamma subunit of bovine retinal transducin. Proc Natl Acad Sci U S A. 1984 Nov;81(22):6948–6952. doi: 10.1073/pnas.81.22.6948. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Hurley J. B. Molecular properties of the cGMP cascade of vertebrate photoreceptors. Annu Rev Physiol. 1987;49:793–812. doi: 10.1146/annurev.ph.49.030187.004045. [DOI] [PubMed] [Google Scholar]
  21. Itoh H., Kozasa T., Nagata S., Nakamura S., Katada T., Ui M., Iwai S., Ohtsuka E., Kawasaki H., Suzuki K. Molecular cloning and sequence determination of cDNAs for alpha subunits of the guanine nucleotide-binding proteins Gs, Gi, and Go from rat brain. Proc Natl Acad Sci U S A. 1986 Jun;83(11):3776–3780. doi: 10.1073/pnas.83.11.3776. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Jelsema C. L., Axelrod J. Stimulation of phospholipase A2 activity in bovine rod outer segments by the beta gamma subunits of transducin and its inhibition by the alpha subunit. Proc Natl Acad Sci U S A. 1987 Jun;84(11):3623–3627. doi: 10.1073/pnas.84.11.3623. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Katada T., Kusakabe K., Oinuma M., Ui M. A novel mechanism for the inhibition of adenylate cyclase via inhibitory GTP-binding proteins. Calmodulin-dependent inhibition of the cyclase catalyst by the beta gamma-subunits of GTP-binding proteins. J Biol Chem. 1987 Sep 5;262(25):11897–11900. [PubMed] [Google Scholar]
  24. Kimura M. Estimation of evolutionary distances between homologous nucleotide sequences. Proc Natl Acad Sci U S A. 1981 Jan;78(1):454–458. doi: 10.1073/pnas.78.1.454. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Logothetis D. E., Kurachi Y., Galper J., Neer E. J., Clapham D. E. The beta gamma subunits of GTP-binding proteins activate the muscarinic K+ channel in heart. Nature. 1987 Jan 22;325(6102):321–326. doi: 10.1038/325321a0. [DOI] [PubMed] [Google Scholar]
  26. Maniatis T., Hardison R. C., Lacy E., Lauer J., O'Connell C., Quon D., Sim G. K., Efstratiadis A. The isolation of structural genes from libraries of eucaryotic DNA. Cell. 1978 Oct;15(2):687–701. doi: 10.1016/0092-8674(78)90036-3. [DOI] [PubMed] [Google Scholar]
  27. Messing J. New M13 vectors for cloning. Methods Enzymol. 1983;101:20–78. doi: 10.1016/0076-6879(83)01005-8. [DOI] [PubMed] [Google Scholar]
  28. Mumby S. M., Kahn R. A., Manning D. R., Gilman A. G. Antisera of designed specificity for subunits of guanine nucleotide-binding regulatory proteins. Proc Natl Acad Sci U S A. 1986 Jan;83(2):265–269. doi: 10.1073/pnas.83.2.265. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Munn T. Z., Mues G. I. Human lipocortin similar to ras gene products. Nature. 1986 Jul 24;322(6077):314–315. doi: 10.1038/322314a0. [DOI] [PubMed] [Google Scholar]
  30. Nakafuku M., Itoh H., Nakamura S., Kaziro Y. Occurrence in Saccharomyces cerevisiae of a gene homologous to the cDNA coding for the alpha subunit of mammalian G proteins. Proc Natl Acad Sci U S A. 1987 Apr;84(8):2140–2144. doi: 10.1073/pnas.84.8.2140. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. O'Connell P. O., Rosbash M. Sequence, structure, and codon preference of the Drosophila ribosomal protein 49 gene. Nucleic Acids Res. 1984 Jul 11;12(13):5495–5513. doi: 10.1093/nar/12.13.5495. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Poole S. J., Kauvar L. M., Drees B., Kornberg T. The engrailed locus of Drosophila: structural analysis of an embryonic transcript. Cell. 1985 Jan;40(1):37–43. doi: 10.1016/0092-8674(85)90306-x. [DOI] [PubMed] [Google Scholar]
  33. 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]
  34. Saris C. J., Tack B. F., Kristensen T., Glenney J. R., Jr, Hunter T. The cDNA sequence for the protein-tyrosine kinase substrate p36 (calpactin I heavy chain) reveals a multidomain protein with internal repeats. Cell. 1986 Jul 18;46(2):201–212. doi: 10.1016/0092-8674(86)90737-3. [DOI] [PubMed] [Google Scholar]
  35. Sugimoto K., Nukada T., Tanabe T., Takahashi H., Noda M., Minamino N., Kangawa K., Matsuo H., Hirose T., Inayama S. Primary structure of the beta-subunit of bovine transducin deduced from the cDNA sequence. FEBS Lett. 1985 Oct 28;191(2):235–240. doi: 10.1016/0014-5793(85)80015-6. [DOI] [PubMed] [Google Scholar]
  36. Yoshioka T., Inoue H., Takagi M., Hayashi F., Amakawa T. The effect of isobutylmethylxanthine on the photoresponse and phosphorylation of phosphatidylinositol in octopus retina. Biochim Biophys Acta. 1983 Jan 4;755(1):50–55. doi: 10.1016/0304-4165(83)90271-4. [DOI] [PubMed] [Google Scholar]

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