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 Jul 1;88(13):5587–5591. doi: 10.1073/pnas.88.13.5587

G alpha 16, a G protein alpha subunit specifically expressed in hematopoietic cells.

T T Amatruda 3rd 1, D A Steele 1, V Z Slepak 1, M I Simon 1
PMCID: PMC51922  PMID: 1905813

Abstract

Signal-transduction pathways mediated by guanine nucleotide-binding regulatory proteins (G proteins) determine many of the responses of hematopoietic cells. A recently identified gene encoding a G protein alpha subunit, G alpha 16, is specifically expressed in human cells of the hematopoietic lineage. The G alpha 16 cDNA encodes a protein with predicted Mr of 43,500, which resembles the G q class of alpha subunits and does not include a pertussis toxin ADP-ribosylation site. In comparison with other G protein alpha subunits, the G alpha 16 predicted protein has distinctive amino acid sequences in the amino terminus, the region A guanine nucleotide-binding domain, and in the carboxyl-terminal third of the protein. Cell lines of myelomonocytic and T-cell phenotype express the G alpha 16 gene, but no expression is detectable in two B-cell lines or in nonhematopoietic cell lines. G alpha 16 gene expression is down-regulated in HL-60 cells induced to differentiate to neutrophils with dimethyl sulfoxide. Antisera generated from synthetic peptides that correspond to two regions of G alpha 16 specifically react with a protein of 42- to 43-kDa in bacterial strains that overexpress G alpha 16 and in HL-60 membranes. This protein is decreased in membranes from dimethyl sulfoxide-differentiated HL-60 cells and is not detectable in COS cell membranes. The restricted expression of this gene suggests that G alpha 16 regulates cell-type-specific signal-transduction pathways, which are not inhibited by pertussis toxin.

Full text

PDF
5587

Images in this article

5589Image
on p.5589
5590Image
on p.5590
5590Image
on p.5590
5590Image
on p.5590

Selected References

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

  1. Amatruda T. T., 3rd, Gautam N., Fong H. K., Northup J. K., Simon M. I. The 35- and 36-kDa beta subunits of GTP-binding regulatory proteins are products of separate genes. J Biol Chem. 1988 Apr 15;263(11):5008–5011. [PubMed] [Google Scholar]
  2. Aussel C., Mary D., Peyron J. F., Pelassy C., Ferrua B., Fehlmann M. Inhibition and activation of interleukin 2 synthesis by direct modification of guanosine triphosphate-binding proteins. J Immunol. 1988 Jan 1;140(1):215–220. [PubMed] [Google Scholar]
  3. Beals C. R., Wilson C. B., Perlmutter R. M. A small multigene family encodes Gi signal-transduction proteins. Proc Natl Acad Sci U S A. 1987 Nov;84(22):7886–7890. doi: 10.1073/pnas.84.22.7886. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Bengtsson T., Särndahl E., Stendahl O., Andersson T. Involvement of GTP-binding proteins in actin polymerization in human neutrophils. Proc Natl Acad Sci U S A. 1990 Apr;87(8):2921–2925. doi: 10.1073/pnas.87.8.2921. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Birnbaumer L. G proteins in signal transduction. Annu Rev Pharmacol Toxicol. 1990;30:675–705. doi: 10.1146/annurev.pa.30.040190.003331. [DOI] [PubMed] [Google Scholar]
  6. Bourne H. R., Sanders D. A., McCormick F. The GTPase superfamily: conserved structure and molecular mechanism. Nature. 1991 Jan 10;349(6305):117–127. doi: 10.1038/349117a0. [DOI] [PubMed] [Google Scholar]
  7. 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]
  8. 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]
  9. Chirgwin J. M., Przybyla A. E., MacDonald R. J., Rutter W. J. Isolation of biologically active ribonucleic acid from sources enriched in ribonuclease. Biochemistry. 1979 Nov 27;18(24):5294–5299. doi: 10.1021/bi00591a005. [DOI] [PubMed] [Google Scholar]
  10. Church G. M., Gilbert W. Genomic sequencing. Proc Natl Acad Sci U S A. 1984 Apr;81(7):1991–1995. doi: 10.1073/pnas.81.7.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Collins S. J., Gallo R. C., Gallagher R. E. Continuous growth and differentiation of human myeloid leukaemic cells in suspension culture. Nature. 1977 Nov 24;270(5635):347–349. doi: 10.1038/270347a0. [DOI] [PubMed] [Google Scholar]
  12. Collins S. J., Ruscetti F. W., Gallagher R. E., Gallo R. C. Terminal differentiation of human promyelocytic leukemia cells induced by dimethyl sulfoxide and other polar compounds. Proc Natl Acad Sci U S A. 1978 May;75(5):2458–2462. doi: 10.1073/pnas.75.5.2458. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Didsbury J. R., Ho Y. S., Snyderman R. Human Gi protein alpha-subunit: deduction of amino acid structure from a cloned cDNA. FEBS Lett. 1987 Jan 26;211(2):160–164. doi: 10.1016/0014-5793(87)81428-x. [DOI] [PubMed] [Google Scholar]
  14. Dubyak G. R., Cowen D. S., Meuller L. M. Activation of inositol phospholipid breakdown in HL60 cells by P2-purinergic receptors for extracellular ATP. Evidence for mediation by both pertussis toxin-sensitive and pertussis toxin-insensitive mechanisms. J Biol Chem. 1988 Dec 5;263(34):18108–18117. [PubMed] [Google Scholar]
  15. 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]
  16. Fels A. O., Cohn Z. A. The alveolar macrophage. J Appl Physiol (1985) 1986 Feb;60(2):353–369. doi: 10.1152/jappl.1986.60.2.353. [DOI] [PubMed] [Google Scholar]
  17. Fällman M., Lew D. P., Stendahl O., Andersson T. Receptor-mediated phagocytosis in human neutrophils is associated with increased formation of inositol phosphates and diacylglycerol. Elevation in cytosolic free calcium and formation of inositol phosphates can be dissociated from accumulation of diacylglycerol. J Clin Invest. 1989 Sep;84(3):886–891. doi: 10.1172/JCI114249. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. 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]
  19. Goldman D. W., Chang F. H., Gifford L. A., Goetzl E. J., Bourne H. R. Pertussis toxin inhibition of chemotactic factor-induced calcium mobilization and function in human polymorphonuclear leukocytes. J Exp Med. 1985 Jul 1;162(1):145–156. doi: 10.1084/jem.162.1.145. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Goldsmith P., Gierschik P., Milligan G., Unson C. G., Vinitsky R., Malech H. L., Spiegel A. M. Antibodies directed against synthetic peptides distinguish between GTP-binding proteins in neutrophil and brain. J Biol Chem. 1987 Oct 25;262(30):14683–14688. [PubMed] [Google Scholar]
  21. 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]
  22. 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]
  23. Koeffler H. P., Billing R., Lusis A. J., Sparkes R., Golde D. W. An undifferentiated variant derived from the human acute myelogenous leukemia cell line (KG-1). Blood. 1980 Aug;56(2):265–273. [PubMed] [Google Scholar]
  24. Koeffler H. P., Golde D. W. Acute myelogenous leukemia: a human cell line responsive to colony-stimulating activity. Science. 1978 Jun 9;200(4346):1153–1154. doi: 10.1126/science.306682. [DOI] [PubMed] [Google Scholar]
  25. Kozak M. Point mutations define a sequence flanking the AUG initiator codon that modulates translation by eukaryotic ribosomes. Cell. 1986 Jan 31;44(2):283–292. doi: 10.1016/0092-8674(86)90762-2. [DOI] [PubMed] [Google Scholar]
  26. Krause K. H., Schlegel W., Wollheim C. B., Andersson T., Waldvogel F. A., Lew P. D. Chemotactic peptide activation of human neutrophils and HL-60 cells. Pertussis toxin reveals correlation between inositol trisphosphate generation, calcium ion transients, and cellular activation. J Clin Invest. 1985 Oct;76(4):1348–1354. doi: 10.1172/JCI112109. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Kubonishi I., Machida K., Sonobe H., Ohtsuki Y., Akagi T., Miyoshi I. Two new human myeloid cell lines derived from acute promyelocytic leukemia and chronic myelocytic leukemia. Gan. 1983 Jun;74(3):319–322. [PubMed] [Google Scholar]
  28. Lackie J. M. The behavioural repertoire of neutrophils requires multiple signal transduction pathways. J Cell Sci. 1988 Apr;89(Pt 4):449–452. doi: 10.1242/jcs.89.4.449. [DOI] [PubMed] [Google Scholar]
  29. Linder M. E., Ewald D. A., Miller R. J., Gilman A. G. Purification and characterization of Go alpha and three types of Gi alpha after expression in Escherichia coli. J Biol Chem. 1990 May 15;265(14):8243–8251. [PubMed] [Google Scholar]
  30. 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]
  31. Lozzio C. B., Lozzio B. B. Human chronic myelogenous leukemia cell-line with positive Philadelphia chromosome. Blood. 1975 Mar;45(3):321–334. [PubMed] [Google Scholar]
  32. Luetje C. W., Gierschik P., Milligan G., Unson C., Spiegel A., Nathanson N. M. Tissue-specific regulation of GTP-binding protein and muscarinic acetylcholine receptor levels during cardiac development. Biochemistry. 1987 Jul 28;26(15):4876–4884. doi: 10.1021/bi00389a040. [DOI] [PubMed] [Google Scholar]
  33. Martin P., Papayannopoulou T. HEL cells: a new human erythroleukemia cell line with spontaneous and induced globin expression. Science. 1982 Jun 11;216(4551):1233–1235. doi: 10.1126/science.6177045. [DOI] [PubMed] [Google Scholar]
  34. Masters S. B., Stroud R. M., Bourne H. R. Family of G protein alpha chains: amphipathic analysis and predicted structure of functional domains. Protein Eng. 1986 Oct-Nov;1(1):47–54. [PubMed] [Google Scholar]
  35. 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]
  36. Pang I. H., Sternweis P. C. Purification of unique alpha subunits of GTP-binding regulatory proteins (G proteins) by affinity chromatography with immobilized beta gamma subunits. J Biol Chem. 1990 Oct 25;265(30):18707–18712. [PubMed] [Google Scholar]
  37. Ponte P., Gunning P., Blau H., Kedes L. Human actin genes are single copy for alpha-skeletal and alpha-cardiac actin but multicopy for beta- and gamma-cytoskeletal genes: 3' untranslated regions are isotype specific but are conserved in evolution. Mol Cell Biol. 1983 Oct;3(10):1783–1791. doi: 10.1128/mcb.3.10.1783. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. 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]
  39. Shackleford G. M., Varmus H. E. Expression of the proto-oncogene int-1 is restricted to postmeiotic male germ cells and the neural tube of mid-gestational embryos. Cell. 1987 Jul 3;50(1):89–95. doi: 10.1016/0092-8674(87)90665-9. [DOI] [PubMed] [Google Scholar]
  40. Simon M. I., Strathmann M. P., Gautam N. Diversity of G proteins in signal transduction. Science. 1991 May 10;252(5007):802–808. doi: 10.1126/science.1902986. [DOI] [PubMed] [Google Scholar]
  41. Smith C. D., Lane B. C., Kusaka I., Verghese M. W., Snyderman R. Chemoattractant receptor-induced hydrolysis of phosphatidylinositol 4,5-bisphosphate in human polymorphonuclear leukocyte membranes. Requirement for a guanine nucleotide regulatory protein. J Biol Chem. 1985 May 25;260(10):5875–5878. [PubMed] [Google Scholar]
  42. 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]
  43. 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]
  44. 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]
  45. Stryer L., Bourne H. R. G proteins: a family of signal transducers. Annu Rev Cell Biol. 1986;2:391–419. doi: 10.1146/annurev.cb.02.110186.002135. [DOI] [PubMed] [Google Scholar]
  46. Studier F. W., Moffatt B. A. Use of bacteriophage T7 RNA polymerase to direct selective high-level expression of cloned genes. J Mol Biol. 1986 May 5;189(1):113–130. doi: 10.1016/0022-2836(86)90385-2. [DOI] [PubMed] [Google Scholar]
  47. Tsuchiya S., Kobayashi Y., Goto Y., Okumura H., Nakae S., Konno T., Tada K. Induction of maturation in cultured human monocytic leukemia cells by a phorbol diester. Cancer Res. 1982 Apr;42(4):1530–1536. [PubMed] [Google Scholar]
  48. Verghese M. W., Smith C. D., Charles L. A., Jakoi L., Synderman R. A guanine nucleotide regulatory protein controls polyphosphoinositide metabolism, Ca2+ mobilization, and cellular responses to chemoattractants in human monocytes. J Immunol. 1986 Jul 1;137(1):271–275. [PubMed] [Google Scholar]
  49. Watkins D. C., Northup J. K., Malbon C. C. Regulation of G-proteins in differentiation. Altered ratio of alpha- to beta-subunits in 3T3-L1 cells. J Biol Chem. 1987 Aug 5;262(22):10651–10657. [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