Molecular and Cellular Biochemistry

, Volume 144, Issue 1, pp 45–51 | Cite as

Expression of GTP-binding protein alpha subunits in human thymocytes

  • Panagiotis S. Kabouridis
  • Samuel T. Waters
  • Sabine Escobar
  • Jack Stanners
  • Constantine D. Tsoukas


In this report, we investigate G protein α subunit diversity in human thymocytes, utilizing common properties shared by these genes and reverse transcription-polymerase chain reaction (RT-PCR). Sequence analysis of PCR amplified gene portions, indicate the presence of members from all four G-protein families that have been described thus far. The α subunit genes identified are: Gαi1-3 and Gαz but not Gαo from the Gi family, Gαs from the Gs family, Gα11, Gαq, and Gα16 from the Gq family, and Gα12 and Gα13 from the G12 family. Also in this report we present the nucleotide and predicted amino acid sequences of the human Gα13 cloned from a thymocyte cDNA library. The sequence of the human Gα13 has not been previously reported. Comparison of this sequence with the reported murine Gα13 shows >90% identity at the deduced amino acid sequence level. We conclude that thymocytes represent a useful experimental system fo the study of G protein involvement in immune responses and lymphocyte development.

Key words

G proteins thymocytes PCR 


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  1. 1.
    Weiss A, Imboden J, Hardy K, Manger B, Terhorst C, Stobo J: The role of the T3/antigen receptor complex in T-cell activation. Ann Rev Immunol 4: 593–619, 1986Google Scholar
  2. 2.
    Weiss A: T cell antigen receptor signal transduction: A tale of tails and cytoplasmic protein-tyrosine kinases. Cell 73: 209–212, 1993PubMedGoogle Scholar
  3. 3.
    Gilman AG: G proteins: Transducers of receptor-generated signals. Ann Rev Biochem 56: 615–649, 1987PubMedGoogle Scholar
  4. 4.
    Hepler JR, Gilman AG: G proteins. Trends Biochem Sci 17: 383–387, 1992PubMedGoogle Scholar
  5. 5.
    Grunwald GB, Gierschik P, Nirenberg M, Spiegel A: Detection of α-Transducin in retinal rods but not cones. Science 231: 856–859, 1986PubMedGoogle Scholar
  6. 6.
    Jones DT, Reed RR: Golf: An olfactory neuron specific-G protein involved in odorant signal transduction. Science 244: 790–795, 1989PubMedGoogle Scholar
  7. 7.
    McLaughlin SK, McKinnon PJ, Margolskee RF: Gustducin is a tastecell-specific G protein closely related to the transducins. Nature 357: 563–569, 1992PubMedGoogle Scholar
  8. 8.
    Imboden JB, Stobo JD: Transmembrane signalling by the T cell antigen receptor. Perturbation of the T3-antigen receptor complex generates inositol phosphates and releases calcium ions from intracellular stores. J Exp Med 161: 446–456, 1985PubMedGoogle Scholar
  9. 9.
    Meisenhelder J, Suh PG, Rhee SG, Hunter T: Phospholipase C-γ is a substrate for the PDGF and EGF receptor protein tyrosine kinasesin vivo andin vitro. Cell 57: 1109–1122, 1989PubMedGoogle Scholar
  10. 10.
    Anderson DL, Tsoukas CD: Cholera toxin inhibits resting human T cell activation via a cAMP-independent pathway. J Immunol 143: 3647–3652, 1989PubMedGoogle Scholar
  11. 11.
    Imboden JB, Shoback DM, Pattison G, Stobo JD: Cholera toxin inhibits the T-cell antigen receptor-mediated increases in inositol triphosphate and cytoplasmic free calcium. Proc Natl Acad Sci, USA 83: 5673–5677, 1986Google Scholar
  12. 12.
    O'Shea JJ, Urdahl KB, Luong HT, Chused TM, Samuelson LE, Klausner RD: Aluminium fluoride induces phosphatidylinositol turnover, elevation of cytoplasmic free calcium, and phosphorylation of the T-cell antigen receptor in murine T-cells. J Immunol 139: 3463–3469, 1987PubMedGoogle Scholar
  13. 13.
    Graves JD, Cantrell DA. An Analysis of the Role of Nucleotide Binding Proteins in Antigen Receptor/CD3 Antigen Coupling to Phospholipase C. J Immunol 146: 2102–2107, 1991PubMedGoogle Scholar
  14. 14.
    Huang C-L and Ives, HE: Guanosine 5′-O-(3-thiotrisphosphate) potentiates both thrombin- and platelet-derived growth factor-induced inositol phosphate release in permiabilized vascular smooth muscle cells. J Biol Chem 264: 4391–4397, 1989PubMedGoogle Scholar
  15. 15.
    Moingeon P, Jin Y-J, Stebbins CC, Lopez PA, Alcover A, Reinherz EL: Characterization of Functional GTP Binding Proteins in Jurkat T Cell Mutants Lacking Either CD3-Ti or CD2 Surface Receptors. Cell Immunol 128: 578–588, 1990PubMedGoogle Scholar
  16. 16.
    Strathmann M, Wilkie TM, Simon MI: Diversity of the G-protein family: Sequences from five additional alpha subunits in the mouse. Proc Natl Acad Sci USA 86: 7407–7409, 1989PubMedGoogle Scholar
  17. 17.
    Chomczynski P and Sacchi N: Single-step method of RNA isolation by acid guanidinium-thiocyanate-phenol-chloroform extraction. Anal Biochem 162: 156–159, 1987PubMedGoogle Scholar
  18. 18.
    Berridge MJ, Irvine RF: Inositol phosphates and cell signalling. Nature 341: 197–205, 1989PubMedGoogle Scholar
  19. 19.
    Weiss A, Koretzky G, Schatzman RC, Kadlecek T: Functional activation of the T-cell antigen receptor induces tyrosine phosphorylation of phospholipase C-γl. Proc Natl Acad Sci USA 88: 5484–5488, 1991PubMedGoogle Scholar
  20. 20.
    Chaffin KE, Perlmutter RM: A pertussis toxin-sensitive process controls thymocyte emigration. Eur J Immunol 21: 2565–2573, 1991PubMedGoogle Scholar
  21. 21.
    Schrezenmeier H, Ahnert-Hilger G, Fleischer B: T cell receptor-associated GTP-binding triggers T cell receptor-mediated granule exocytosis in cytotoxic T lymphocytes. J Immunol 141: 3785–3790, 1988PubMedGoogle Scholar
  22. 22.
    Schrezenmeier H, Anhert-Hilger G, Fleischer B: Inactivation of a T cell receptor-associated GTP-binding protein by antibody-induced modulation of the T cell receptor/CD3 complex. J Exp Med 178: 817–822, 1988Google Scholar
  23. 23.
    Cenciarelli C, Hohman RJ, Atkinson TP, Gusovsky F, Weissman AM: Evidence for GTP-binding Protein Involvement in the Tyrosine Phosphorylation of the T Cell Receptor Zeta Chain. J Biol Chem 267: 14527–14530, 1992PubMedGoogle Scholar
  24. 24.
    Simon MI, Strathmann MP, Gautman N: Diversity of G proteins in signal transduction. Science 252: 802–808, 1991PubMedGoogle Scholar
  25. 25.
    Holter W, Spiegel AM, Howard BH, Weber S, Brann MR: Expression of GTP-Binding Proteins and Prostaglandin E2 Receptors during Human T Cell Antigen. Cell Immunol 134: 287–295, 1991PubMedGoogle Scholar
  26. 26.
    Strathmann MP, Simon MI: Gα12 and Gα13 subunits define a fourth class of G protein α subunits. Proc Natl Acad Sci USA 88: 5582–5586, 1991PubMedGoogle Scholar
  27. 27.
    Strathmann MP, Simon MI: G Protein Diversity: A distinct class of alpha subunits is present in vertebrates and invertebrates. Proc Natl Acad Sci USA 87: 9113–9117, 1990PubMedGoogle Scholar
  28. 28.
    Beals CR, Wilson CB, Perlmutter RM A small multigene family encodes Gi signal-transduction proteins. Proc Natl Acad Sci USA 84: 7886–7890, 1987PubMedGoogle Scholar
  29. 29.
    Sullivan KA, Liao YC, Alborzi A, Beiderman B, Chang FH, Masters SB, Levinson AD, Bourne HR: Inhibitory and stimulatory G proteins of adenylate cyclase: cDNA and amino acid sequences of the α chains. Proc Natl Acad Sci USA 83: 6687–6691, 1986PubMedGoogle Scholar
  30. 30.
    Goldsmith MA, Desai DM, Schultz T, Weiss A: Function of a heterologous muscarinic receptor in T cell antigen receptor signal transduction mutants. J Biol Chem 264: 17190–17197, 1989PubMedGoogle Scholar
  31. 31.
    Desai DM, Newton ME, Kadlecek T, Weiss A: Stimulation of the phosphotidyl-inositol pathway can induce T-cell activation. Nature 348: 66–69, 1990PubMedGoogle Scholar
  32. 32.
    Berstein G BJL, Smrcka AV, Higashijima T, Sternweis PC, Exton JH and Ross EM: Reconstitution of agonist-stimulated phosphatidylinositol 4,5-biphosphate hydrolysis using purified m1 muscarinic receptor, Gq/11, and phospholipase C-beta 1. J Biol Chem 267: 8081–8088, 1992PubMedGoogle Scholar

Copyright information

© Kluwer Academic Publishers 1995

Authors and Affiliations

  • Panagiotis S. Kabouridis
    • 1
    • 2
  • Samuel T. Waters
    • 1
    • 2
  • Sabine Escobar
    • 1
    • 2
  • Jack Stanners
    • 1
    • 2
  • Constantine D. Tsoukas
    • 1
    • 3
    • 2
  1. 1.Department of BiologySan Diego State UniversitySan DiegoUSA
  2. 2.Molecular Biology InstituteSan Diego State UniversitySan DiegoUSA
  3. 3.Department of Molecular and Experimental MedicineScripps Research InstituteLa JollaUSA

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