Skip to main content
NCBI home page
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
. 1991 Apr 15;88(8):2974–2978. doi: 10.1073/pnas.88.8.2974

Structure of the human gene and two rat cDNAs encoding the alpha chain of GTP-binding regulatory protein Go: two different mRNAs are generated by alternative splicing.

T Tsukamoto 1, R Toyama 1, H Itoh 1, T Kozasa 1, M Matsuoka 1, Y Kaziro 1
PMCID: PMC51366  PMID: 1901650

Abstract

Go is a specific class ("other") of signal-transducing heterotrimeric GTP-binding proteins (G proteins) that is expressed in high levels in mammalian brain. We have cloned two different rat cDNAs encoding the alpha subunit of Go (Go alpha-1 and Go alpha-2) and a human Go alpha chromosomal gene. The human Go alpha gene spans more than 100 kilobases and contains 11 exons, including one noncoding exon in the 3' flanking region. The 5' flanking region is highly G + C-rich and contains five G.C boxes (Sp1 binding sites) but no TATA box. Exons 7 and 8 coding for amino acid residues 242-354 of Go alpha protein are duplicated (referred to as exons 7A, 7B, 8A, and 8B). It was found that exons 7A and 8A code for Go alpha-1, and 7B and 8B code for Go alpha-2. This indicates that two different Go alpha mRNAs may be generated by alternative splicing of a single Go alpha gene. The splice sites of the Go alpha-1 and Go alpha-2 genes are completely identical with those encoding human inhibitory G protein alpha subunits Gi2 alpha and Gi3 alpha [Itoh, H., Toyama, R., Kozasa, T., Tsukamoto, T., Matsuoka, M. & Kaziro, Y. (1988) J. Biol. Chem. 263, 6656-6664] and also transducin G protein alpha subunit Gt1 alpha [Raport, C. J., Dere, B. & Hurley, J. (1989) J. Biol. Chem. 264, 7122-7128]. Sequence homology and conservation of the exon-intron organization indicate that the genes coding for Go alpha, Gi2 alpha, Gi3 alpha, Gt1 alpha, and probably Gi1 alpha may be evolved from a common progenitor. Like Go alpha-1, Go alpha-2 is expressed mainly in brain.

Full text

PDF
2974

Images in this article

2977Image
on p.2977
2977Image
on p.2977

Selected References

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

  1. Asano T., Semba R., Kamiya N., Ogasawara N., Kato K. Go, a GTP-binding protein: immunochemical and immunohistochemical localization in the rat. J Neurochem. 1988 Apr;50(4):1164–1169. doi: 10.1111/j.1471-4159.1988.tb10588.x. [DOI] [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. Brann M. R., Collins R. M., Spiegel A. Localization of mRNAs encoding the alpha-subunits of signal-transducing G-proteins within rat brain and among peripheral tissues. FEBS Lett. 1987 Sep 28;222(1):191–198. doi: 10.1016/0014-5793(87)80218-1. [DOI] [PubMed] [Google Scholar]
  4. Bray P., Carter A., Simons C., Guo V., Puckett C., Kamholz J., Spiegel A., Nirenberg M. Human cDNA clones for four species of G alpha s signal transduction protein. Proc Natl Acad Sci U S A. 1986 Dec;83(23):8893–8897. doi: 10.1073/pnas.83.23.8893. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Feinberg A. P., Vogelstein B. A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity. Anal Biochem. 1983 Jul 1;132(1):6–13. doi: 10.1016/0003-2697(83)90418-9. [DOI] [PubMed] [Google Scholar]
  6. Fong H. K., Yoshimoto K. K., Eversole-Cire P., Simon M. I. Identification of a GTP-binding protein alpha subunit that lacks an apparent ADP-ribosylation site for pertussis toxin. Proc Natl Acad Sci U S A. 1988 May;85(9):3066–3070. doi: 10.1073/pnas.85.9.3066. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Freissmuth M., Casey P. J., Gilman A. G. G proteins control diverse pathways of transmembrane signaling. FASEB J. 1989 Aug;3(10):2125–2131. [PubMed] [Google Scholar]
  8. 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]
  9. Goldsmith P., Backlund P. S., Jr, Rossiter K., Carter A., Milligan G., Unson C. G., Spiegel A. Purification of heterotrimeric GTP-binding proteins from brain: identification of a novel form of Go. Biochemistry. 1988 Sep 6;27(18):7085–7090. doi: 10.1021/bi00418a062. [DOI] [PubMed] [Google Scholar]
  10. Hsu W. H., Rudolph U., Sanford J., Bertrand P., Olate J., Nelson C., Moss L. G., Boyd A. E., Codina J., Birnbaumer L. Molecular cloning of a novel splice variant of the alpha subunit of the mammalian Go protein. J Biol Chem. 1990 Jul 5;265(19):11220–11226. [PubMed] [Google Scholar]
  11. Huff R. M., Axton J. M., Neer E. J. Physical and immunological characterization of a guanine nucleotide-binding protein purified from bovine cerebral cortex. J Biol Chem. 1985 Sep 5;260(19):10864–10871. [PubMed] [Google Scholar]
  12. Ishii S., Xu Y. H., Stratton R. H., Roe B. A., Merlino G. T., Pastan I. Characterization and sequence of the promoter region of the human epidermal growth factor receptor gene. Proc Natl Acad Sci U S A. 1985 Aug;82(15):4920–4924. doi: 10.1073/pnas.82.15.4920. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. 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]
  14. Itoh H., Toyama R., Kozasa T., Tsukamoto T., Matsuoka M., Kaziro Y. Presence of three distinct molecular species of Gi protein alpha subunit. Structure of rat cDNAs and human genomic DNAs. J Biol Chem. 1988 May 15;263(14):6656–6664. [PubMed] [Google Scholar]
  15. Jones D. T., Reed R. R. Golf: an olfactory neuron specific-G protein involved in odorant signal transduction. Science. 1989 May 19;244(4906):790–795. doi: 10.1126/science.2499043. [DOI] [PubMed] [Google Scholar]
  16. Jones D. T., Reed R. R. Molecular cloning of five GTP-binding protein cDNA species from rat olfactory neuroepithelium. J Biol Chem. 1987 Oct 15;262(29):14241–14249. [PubMed] [Google Scholar]
  17. Kaziro Y., Itoh H., Kozasa T., Toyama R., Tsukamoto T., Matsuoka M., Nakafuku M., Obara T., Takagi T., Hernandez R. Structures of the genes coding for G-protein alpha subunits from mammalian and yeast cells. Cold Spring Harb Symp Quant Biol. 1988;53(Pt 1):209–220. doi: 10.1101/sqb.1988.053.01.027. [DOI] [PubMed] [Google Scholar]
  18. Kozasa T., Itoh H., Tsukamoto T., Kaziro Y. Isolation and characterization of the human Gs alpha gene. Proc Natl Acad Sci U S A. 1988 Apr;85(7):2081–2085. doi: 10.1073/pnas.85.7.2081. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Lavu S., Clark J., Swarup R., Matsushima K., Paturu K., Moss J., Kung H. F. Molecular cloning and DNA sequence analysis of the human guanine nucleotide-binding protein Go alpha. Biochem Biophys Res Commun. 1988 Jan 29;150(2):811–815. doi: 10.1016/0006-291x(88)90463-9. [DOI] [PubMed] [Google Scholar]
  20. Lawn R. M., Fritsch E. F., Parker R. C., Blake G., Maniatis T. The isolation and characterization of linked delta- and beta-globin genes from a cloned library of human DNA. Cell. 1978 Dec;15(4):1157–1174. doi: 10.1016/0092-8674(78)90043-0. [DOI] [PubMed] [Google Scholar]
  21. Masters S. B., Sullivan K. A., Miller R. T., Beiderman B., Lopez N. G., Ramachandran J., Bourne H. R. Carboxyl terminal domain of Gs alpha specifies coupling of receptors to stimulation of adenylyl cyclase. Science. 1988 Jul 22;241(4864):448–451. doi: 10.1126/science.2899356. [DOI] [PubMed] [Google Scholar]
  22. Matsuoka M., Itoh H., Kozasa T., Kaziro Y. Sequence analysis of cDNA and genomic DNA for a putative pertussis toxin-insensitive guanine nucleotide-binding regulatory protein alpha subunit. Proc Natl Acad Sci U S A. 1988 Aug;85(15):5384–5388. doi: 10.1073/pnas.85.15.5384. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Matsushime H., Wang L. H., Shibuya M. Human c-ros-1 gene homologous to the v-ros sequence of UR2 sarcoma virus encodes for a transmembrane receptorlike molecule. Mol Cell Biol. 1986 Aug;6(8):3000–3004. doi: 10.1128/mcb.6.8.3000. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Mattera R., Codina J., Crozat A., Kidd V., Woo S. L., Birnbaumer L. Identification by molecular cloning of two forms of the alpha-subunit of the human liver stimulatory (GS) regulatory component of adenylyl cyclase. FEBS Lett. 1986 Sep 29;206(1):36–42. doi: 10.1016/0014-5793(86)81336-9. [DOI] [PubMed] [Google Scholar]
  25. 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]
  26. Milligan G., Tanfin Z., Goureau O., Unson C., Harbon S. Identification of both Gi2 and a novel, immunologically distinct, form of Go in rat myometrial membranes. FEBS Lett. 1989 Feb 27;244(2):411–416. doi: 10.1016/0014-5793(89)80574-5. [DOI] [PubMed] [Google Scholar]
  27. Mizusawa S., Nishimura S., Seela F. Improvement of the dideoxy chain termination method of DNA sequencing by use of deoxy-7-deazaguanosine triphosphate in place of dGTP. Nucleic Acids Res. 1986 Feb 11;14(3):1319–1324. doi: 10.1093/nar/14.3.1319. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Mullaney I., Milligan G. Elevated levels of the guanine nucleotide binding protein, Go, are associated with differentiation of neuroblastoma x glioma hybrid cells. FEBS Lett. 1989 Feb 13;244(1):113–118. doi: 10.1016/0014-5793(89)81174-3. [DOI] [PubMed] [Google Scholar]
  29. 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]
  30. Neer E. J., Clapham D. E. Roles of G protein subunits in transmembrane signalling. Nature. 1988 May 12;333(6169):129–134. doi: 10.1038/333129a0. [DOI] [PubMed] [Google Scholar]
  31. Neer E. J., Lok J. M., Wolf L. G. Purification and properties of the inhibitory guanine nucleotide regulatory unit of brain adenylate cyclase. J Biol Chem. 1984 Nov 25;259(22):14222–14229. [PubMed] [Google Scholar]
  32. Olate J., Jorquera H., Purcell P., Codina J., Birnbaumer L., Allende J. E. Molecular cloning and sequence determination of a cDNA coding for the alpha-subunit of a Go-type protein of Xenopus laevis oocytes. FEBS Lett. 1989 Feb 13;244(1):188–192. doi: 10.1016/0014-5793(89)81190-1. [DOI] [PubMed] [Google Scholar]
  33. Ovchinnikov YuA, Slepak V. Z., Pronin A. N., Shlensky A. B., Levina N. B., Voeikov V. L., Lipkin V. M. Primary structure of bovine cerebellum GTP-binding protein G39 and its effect on the adenylate cyclase system. FEBS Lett. 1987 Dec 21;226(1):91–95. doi: 10.1016/0014-5793(87)80557-4. [DOI] [PubMed] [Google Scholar]
  34. Price S. R., Murtagh J. J., Jr, Tsuchiya M., Serventi I. M., Van Meurs K. P., Angus C. W., Moss J., Vaughan M. Multiple forms of Go alpha mRNA: analysis of the 3'-untranslated regions. Biochemistry. 1990 May 29;29(21):5069–5076. doi: 10.1021/bi00473a011. [DOI] [PubMed] [Google Scholar]
  35. Price S. R., Tsai S. C., Adamik R., Angus C. W., Serventi I. M., Tsuchiya M., Moss J., Vaughan M. Expression of Go alpha mRNA and protein in bovine tissues. Biochemistry. 1989 May 2;28(9):3803–3807. doi: 10.1021/bi00435a027. [DOI] [PubMed] [Google Scholar]
  36. Raport C. J., Dere B., Hurley J. B. Characterization of the mouse rod transducin alpha subunit gene. J Biol Chem. 1989 May 5;264(13):7122–7128. [PubMed] [Google Scholar]
  37. 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]
  38. Robishaw J. D., Smigel M. D., Gilman A. G. Molecular basis for two forms of the G protein that stimulates adenylate cyclase. J Biol Chem. 1986 Jul 25;261(21):9587–9590. [PubMed] [Google Scholar]
  39. 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]
  40. Sternweis P. C., Robishaw J. D. Isolation of two proteins with high affinity for guanine nucleotides from membranes of bovine brain. J Biol Chem. 1984 Nov 25;259(22):13806–13813. [PubMed] [Google Scholar]
  41. Strathmann M., Wilkie T. M., Simon M. I. Alternative splicing produces transcripts encoding two forms of the alpha subunit of GTP-binding protein Go. Proc Natl Acad Sci U S A. 1990 Sep;87(17):6477–6481. doi: 10.1073/pnas.87.17.6477. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Thambi N. C., Quan F., Wolfgang W. J., Spiegel A., Forte M. Immunological and molecular characterization of Go alpha-like proteins in the Drosophila central nervous system. J Biol Chem. 1989 Nov 5;264(31):18552–18560. [PubMed] [Google Scholar]
  43. Van Meurs K. P., Angus C. W., Lavu S., Kung H. F., Czarnecki S. K., Moss J., Vaughan M. Deduced amino acid sequence of bovine retinal Go alpha: similarities to other guanine nucleotide-binding proteins. Proc Natl Acad Sci U S A. 1987 May;84(10):3107–3111. doi: 10.1073/pnas.84.10.3107. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. Yoon J., Shortridge R. D., Bloomquist B. T., Schneuwly S., Perdew M. H., Pak W. L. Molecular characterization of Drosophila gene encoding G0 alpha subunit homolog. J Biol Chem. 1989 Nov 5;264(31):18536–18543. [PubMed] [Google Scholar]
  45. de Sousa S. M., Hoveland L. L., Yarfitz S., Hurley J. B. The Drosophila Go alpha-like G protein gene produces multiple transcripts and is expressed in the nervous system and in ovaries. J Biol Chem. 1989 Nov 5;264(31):18544–18551. [PubMed] [Google Scholar]

Articles from Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences

RESOURCES