Skip to main content
Cell Regulation logoLink to Cell Regulation
. 1991 Feb;2(2):135–154. doi: 10.1091/mbc.2.2.135

Homologous and unique G protein alpha subunits in the nematode Caenorhabditis elegans.

M A Lochrie 1, J E Mendel 1, P W Sternberg 1, M I Simon 1
PMCID: PMC361731  PMID: 1907494

Abstract

A cDNA corresponding to a known G protein alpha subunit, the alpha subunit of Go (Go alpha), was isolated and sequenced. The predicted amino acid sequence of C. elegans Go alpha is 80-87% identical to other Go alpha sequences. An mRNA that hybridizes to the C. elegans Go alpha cDNA can be detected on Northern blots. A C. elegans protein that crossreacts with antibovine Go alpha antibody can be detected on immunoblots. A cosmid clone containing the C. elegans Go alpha gene (goa-1) was isolated and mapped to chromosome I. The genomic fragments of three other C. elegans G protein alpha subunit genes (gpa-1, gpa-2, and gpa-3) have been isolated using the polymerase chain reaction. The corresponding cosmid clones were isolated and mapped to disperse locations on chromosome V. The sequences of two of the genes, gpa-1 and gpa-3, were determined. The predicted amino acid sequences of gpa-1 and gpa-3 are only 48% identical to each other. Therefore, they are likely to have distinct functions. In addition they are not homologous enough to G protein alpha subunits in other organisms to be classified. Thus C. elegans has G proteins that are identifiable homologues of mammalian G proteins as well as G proteins that appear to be unique to C. elegans. Study of identifiable G proteins in C. elegans may result in a further understanding of their function in other organisms, whereas study of the novel G proteins may provide an understanding of unique aspects of nematode physiology.

Full text

PDF
135

Images in this article

Selected References

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

  1. Ballinger D. G., Benzer S. Targeted gene mutations in Drosophila. Proc Natl Acad Sci U S A. 1989 Dec;86(23):9402–9406. doi: 10.1073/pnas.86.23.9402. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. 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]
  3. Brenner S. The genetics of Caenorhabditis elegans. Genetics. 1974 May;77(1):71–94. doi: 10.1093/genetics/77.1.71. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Buss J. E., Mumby S. M., Casey P. J., Gilman A. G., Sefton B. M. Myristoylated alpha subunits of guanine nucleotide-binding regulatory proteins. Proc Natl Acad Sci U S A. 1987 Nov;84(21):7493–7497. doi: 10.1073/pnas.84.21.7493. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Caron M. G. The guanine nucleotide regulatory protein-coupled receptors for nucleosides, nucleotides, amino acids and amine neurotransmitters. Curr Opin Cell Biol. 1989 Apr;1(2):159–166. doi: 10.1016/0955-0674(89)90081-1. [DOI] [PubMed] [Google Scholar]
  6. Casey P. J., Fong H. K., Simon M. I., Gilman A. G. Gz, a guanine nucleotide-binding protein with unique biochemical properties. J Biol Chem. 1990 Feb 5;265(4):2383–2390. [PubMed] [Google Scholar]
  7. Cerione R. A., Regan J. W., Nakata H., Codina J., Benovic J. L., Gierschik P., Somers R. L., Spiegel A. M., Birnbaumer L., Lefkowitz R. J. Functional reconstitution of the alpha 2-adrenergic receptor with guanine nucleotide regulatory proteins in phospholipid vesicles. J Biol Chem. 1986 Mar 15;261(8):3901–3909. [PubMed] [Google Scholar]
  8. Coulson A., Sulston J., Brenner S., Karn J. Toward a physical map of the genome of the nematode Caenorhabditis elegans. Proc Natl Acad Sci U S A. 1986 Oct;83(20):7821–7825. doi: 10.1073/pnas.83.20.7821. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Coulson A., Waterston R., Kiff J., Sulston J., Kohara Y. Genome linking with yeast artificial chromosomes. Nature. 1988 Sep 8;335(6186):184–186. doi: 10.1038/335184a0. [DOI] [PubMed] [Google Scholar]
  10. Devereux J., Haeberli P., Smithies O. A comprehensive set of sequence analysis programs for the VAX. Nucleic Acids Res. 1984 Jan 11;12(1 Pt 1):387–395. doi: 10.1093/nar/12.1part1.387. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Emmons S. W., Rosenzweig B., Hirsh D. Arrangement of repeated sequences in the DNA of the nematode Caenorhabditis elegans. J Mol Biol. 1980 Dec 25;144(4):481–500. doi: 10.1016/0022-2836(80)90333-2. [DOI] [PubMed] [Google Scholar]
  12. 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]
  13. Fino Silva I., Plasterk R. H. Characterization of a G-protein alpha-subunit gene from the nematode Caenorhabditis elegans. J Mol Biol. 1990 Oct 20;215(4):483–487. doi: 10.1016/s0022-2836(05)80160-3. [DOI] [PubMed] [Google Scholar]
  14. Freissmuth M., Gilman A. G. Mutations of GS alpha designed to alter the reactivity of the protein with bacterial toxins. Substitutions at ARG187 result in loss of GTPase activity. J Biol Chem. 1989 Dec 25;264(36):21907–21914. [PubMed] [Google Scholar]
  15. Gabrion J., Brabet P., Nguyen Than Dao B., Homburger V., Dumuis A., Sebben M., Rouot B., Bockaert J. Ultrastructural localization of the GTP-binding protein Go in neurons. Cell Signal. 1989;1(1):107–123. doi: 10.1016/0898-6568(89)90025-9. [DOI] [PubMed] [Google Scholar]
  16. Gillespie P. G., Beavo J. A. Characterization of a bovine cone photoreceptor phosphodiesterase purified by cyclic GMP-sepharose chromatography. J Biol Chem. 1988 Jun 15;263(17):8133–8141. [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. Graziano M. P., Gilman A. G. Synthesis in Escherichia coli of GTPase-deficient mutants of Gs alpha. J Biol Chem. 1989 Sep 15;264(26):15475–15482. [PubMed] [Google Scholar]
  19. Gundersen R. E., Devreotes P. N. In vivo receptor-mediated phosphorylation of a G protein in Dictyostelium. Science. 1990 May 4;248(4955):591–593. doi: 10.1126/science.2110382. [DOI] [PubMed] [Google Scholar]
  20. Harris-Warrick R. M., Hammond C., Paupardin-Tritsch D., Homburger V., Rouot B., Bockaert J., Gerschenfeld H. M. An alpha 40 subunit of a GTP-binding protein immunologically related to Go mediates a dopamine-induced decrease of Ca2+ current in snail neurons. Neuron. 1988 Mar;1(1):27–32. doi: 10.1016/0896-6273(88)90206-1. [DOI] [PubMed] [Google Scholar]
  21. Hescheler J., Rosenthal W., Trautwein W., Schultz G. The GTP-binding protein, Go, regulates neuronal calcium channels. 1987 Jan 29-Feb 4Nature. 325(6103):445–447. doi: 10.1038/325445a0. [DOI] [PubMed] [Google Scholar]
  22. Hodgkin J., Horvitz H. R., Brenner S. Nondisjunction Mutants of the Nematode CAENORHABDITIS ELEGANS. Genetics. 1979 Jan;91(1):67–94. doi: 10.1093/genetics/91.1.67. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Holmes D. S., Quigley M. A rapid boiling method for the preparation of bacterial plasmids. Anal Biochem. 1981 Jun;114(1):193–197. doi: 10.1016/0003-2697(81)90473-5. [DOI] [PubMed] [Google Scholar]
  24. 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]
  25. Huang X. Y., Hirsh D. A second trans-spliced RNA leader sequence in the nematode Caenorhabditis elegans. Proc Natl Acad Sci U S A. 1989 Nov;86(22):8640–8644. doi: 10.1073/pnas.86.22.8640. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. 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]
  27. Jones T. L., Simonds W. F., Merendino J. J., Jr, Brann M. R., Spiegel A. M. Myristoylation of an inhibitory GTP-binding protein alpha subunit is essential for its membrane attachment. Proc Natl Acad Sci U S A. 1990 Jan;87(2):568–572. doi: 10.1073/pnas.87.2.568. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Kaiser K., Goodwin S. F. "Site-selected" transposon mutagenesis of Drosophila. Proc Natl Acad Sci U S A. 1990 Mar;87(5):1686–1690. doi: 10.1073/pnas.87.5.1686. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Kanaho Y., Tsai S. C., Adamik R., Hewlett E. L., Moss J., Vaughan M. Rhodopsin-enhanced GTPase activity of the inhibitory GTP-binding protein of adenylate cyclase. J Biol Chem. 1984 Jun 25;259(12):7378–7381. [PubMed] [Google Scholar]
  30. 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]
  31. Kenyon C. The nematode Caenorhabditis elegans. Science. 1988 Jun 10;240(4858):1448–1453. doi: 10.1126/science.3287621. [DOI] [PubMed] [Google Scholar]
  32. Krause M., Hirsh D. A trans-spliced leader sequence on actin mRNA in C. elegans. Cell. 1987 Jun 19;49(6):753–761. doi: 10.1016/0092-8674(87)90613-1. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. 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]
  34. Landis C. A., Masters S. B., Spada A., Pace A. M., Bourne H. R., Vallar L. GTPase inhibiting mutations activate the alpha chain of Gs and stimulate adenylyl cyclase in human pituitary tumours. Nature. 1989 Aug 31;340(6236):692–696. doi: 10.1038/340692a0. [DOI] [PubMed] [Google Scholar]
  35. Lipman D. J., Pearson W. R. Rapid and sensitive protein similarity searches. Science. 1985 Mar 22;227(4693):1435–1441. doi: 10.1126/science.2983426. [DOI] [PubMed] [Google Scholar]
  36. Lochrie M. A., Simon M. I. G protein multiplicity in eukaryotic signal transduction systems. Biochemistry. 1988 Jul 12;27(14):4957–4965. doi: 10.1021/bi00414a001. [DOI] [PubMed] [Google Scholar]
  37. Masters S. B., Miller R. T., Chi M. H., Chang F. H., Beiderman B., Lopez N. G., Bourne H. R. Mutations in the GTP-binding site of GS alpha alter stimulation of adenylyl cyclase. J Biol Chem. 1989 Sep 15;264(26):15467–15474. [PubMed] [Google Scholar]
  38. McFadzean I., Mullaney I., Brown D. A., Milligan G. Antibodies to the GTP binding protein, Go, antagonize noradrenaline-induced calcium current inhibition in NG108-15 hybrid cells. Neuron. 1989 Aug;3(2):177–182. doi: 10.1016/0896-6273(89)90030-5. [DOI] [PubMed] [Google Scholar]
  39. Milburn M. V., Tong L., deVos A. M., Brünger A., Yamaizumi Z., Nishimura S., Kim S. H. Molecular switch for signal transduction: structural differences between active and inactive forms of protooncogenic ras proteins. Science. 1990 Feb 23;247(4945):939–945. doi: 10.1126/science.2406906. [DOI] [PubMed] [Google Scholar]
  40. Moriarty T. M., Padrell E., Carty D. J., Omri G., Landau E. M., Iyengar R. Go protein as signal transducer in the pertussis toxin-sensitive phosphatidylinositol pathway. Nature. 1990 Jan 4;343(6253):79–82. doi: 10.1038/343079a0. [DOI] [PubMed] [Google Scholar]
  41. Moss J., Vaughan M. ADP-ribosylation of guanyl nucleotide-binding regulatory proteins by bacterial toxins. Adv Enzymol Relat Areas Mol Biol. 1988;61:303–379. doi: 10.1002/9780470123072.ch6. [DOI] [PubMed] [Google Scholar]
  42. Mumby S. M., Heukeroth R. O., Gordon J. I., Gilman A. G. G-protein alpha-subunit expression, myristoylation, and membrane association in COS cells. Proc Natl Acad Sci U S A. 1990 Jan;87(2):728–732. doi: 10.1073/pnas.87.2.728. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. 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]
  44. 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]
  45. Proudfoot N. J., Brownlee G. G. 3' non-coding region sequences in eukaryotic messenger RNA. Nature. 1976 Sep 16;263(5574):211–214. doi: 10.1038/263211a0. [DOI] [PubMed] [Google Scholar]
  46. Provost N. M., Somers D. E., Hurley J. B. A Drosophila melanogaster G protein alpha subunit gene is expressed primarily in embryos and pupae. J Biol Chem. 1988 Aug 25;263(24):12070–12076. [PubMed] [Google Scholar]
  47. Pustell J., Kafatos F. C. A convenient and adaptable package of computer programs for DNA and protein sequence management, analysis and homology determination. Nucleic Acids Res. 1984 Jan 11;12(1 Pt 2):643–655. doi: 10.1093/nar/12.1part2.643. [DOI] [PMC free article] [PubMed] [Google Scholar]
  48. 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]
  49. Sagi-Eisenberg R. GTP-binding proteins as possible targets for protein kinase C action. Trends Biochem Sci. 1989 Sep;14(9):355–357. doi: 10.1016/0968-0004(89)90001-7. [DOI] [PubMed] [Google Scholar]
  50. Schmidt C. J., Garen-Fazio S., Chow Y. K., Neer E. J. Neuronal expression of a newly identified Drosophila melanogaster G protein alpha 0 subunit. Cell Regul. 1989 Nov;1(1):125–134. doi: 10.1091/mbc.1.1.125. [DOI] [PMC free article] [PubMed] [Google Scholar]
  51. Senogles S. E., Spiegel A. M., Padrell E., Iyengar R., Caron M. G. Specificity of receptor-G protein interactions. Discrimination of Gi subtypes by the D2 dopamine receptor in a reconstituted system. J Biol Chem. 1990 Mar 15;265(8):4507–4514. [PubMed] [Google Scholar]
  52. 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]
  53. Strathmann M., Simon M. I. G protein diversity: a distinct class of alpha subunits is present in vertebrates and invertebrates. Proc Natl Acad Sci U S A. 1990 Dec;87(23):9113–9117. doi: 10.1073/pnas.87.23.9113. [DOI] [PMC free article] [PubMed] [Google Scholar]
  54. 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]
  55. Strathmann M., Wilkie T. M., Simon M. I. Diversity of the G-protein family: sequences from five additional alpha subunits in the mouse. Proc Natl Acad Sci U S A. 1989 Oct;86(19):7407–7409. doi: 10.1073/pnas.86.19.7407. [DOI] [PMC free article] [PubMed] [Google Scholar]
  56. Strittmatter S. M., Valenzuela D., Kennedy T. E., Neer E. J., Fishman M. C. G0 is a major growth cone protein subject to regulation by GAP-43. Nature. 1990 Apr 26;344(6269):836–841. doi: 10.1038/344836a0. [DOI] [PubMed] [Google Scholar]
  57. 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]
  58. Towbin H., Staehelin T., Gordon J. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4350–4354. doi: 10.1073/pnas.76.9.4350. [DOI] [PMC free article] [PubMed] [Google Scholar]
  59. Towler D. A., Gordon J. I., Adams S. P., Glaser L. The biology and enzymology of eukaryotic protein acylation. Annu Rev Biochem. 1988;57:69–99. doi: 10.1146/annurev.bi.57.070188.000441. [DOI] [PubMed] [Google Scholar]
  60. 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]
  61. VanDongen A. M., Codina J., Olate J., Mattera R., Joho R., Birnbaumer L., Brown A. M. Newly identified brain potassium channels gated by the guanine nucleotide binding protein Go. Science. 1988 Dec 9;242(4884):1433–1437. doi: 10.1126/science.3144040. [DOI] [PubMed] [Google Scholar]
  62. Weinstein L. S., Spiegel A. M., Carter A. D. Cloning and characterization of the human gene for the alpha-subunit of Gi2, a GTP-binding signal transduction protein. FEBS Lett. 1988 May 23;232(2):333–340. doi: 10.1016/0014-5793(88)80764-6. [DOI] [PubMed] [Google Scholar]
  63. Wickens M., Stephenson P. Role of the conserved AAUAAA sequence: four AAUAAA point mutants prevent messenger RNA 3' end formation. Science. 1984 Nov 30;226(4678):1045–1051. doi: 10.1126/science.6208611. [DOI] [PubMed] [Google Scholar]
  64. Worley P. F., Baraban J. M., Van Dop C., Neer E. J., Snyder S. H. Go, a guanine nucleotide-binding protein: immunohistochemical localization in rat brain resembles distribution of second messenger systems. Proc Natl Acad Sci U S A. 1986 Jun;83(12):4561–4565. doi: 10.1073/pnas.83.12.4561. [DOI] [PMC free article] [PubMed] [Google Scholar]
  65. Yatani A., Mattera R., Codina J., Graf R., Okabe K., Padrell E., Iyengar R., Brown A. M., Birnbaumer L. The G protein-gated atrial K+ channel is stimulated by three distinct Gi alpha-subunits. Nature. 1988 Dec 15;336(6200):680–682. doi: 10.1038/336680a0. [DOI] [PubMed] [Google Scholar]
  66. 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]
  67. 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]
  68. van der Voorn L., Gebbink M., Plasterk R. H., Ploegh H. L. Characterization of a G-protein beta-subunit gene from the nematode Caenorhabditis elegans. J Mol Biol. 1990 May 5;213(1):17–26. doi: 10.1016/s0022-2836(05)80118-4. [DOI] [PubMed] [Google Scholar]

Articles from Cell Regulation are provided here courtesy of American Society for Cell Biology

RESOURCES