Skip to main content
Log in

Expression of guanine nucleotide binding proteins, Gs and Gi, in mRNAs in epidermal keratinocytes

  • Original Contributions
  • Published:
Archives of Dermatological Research Aims and scope Submit manuscript

Summary

The stimulation or inhibition of adenylate cyclase by hormones and chemicals is mediated by the stimulatory or inhibitory guanine nucleotide-binding proteins, Gs and Gi respectively. Although the presence of these G-proteins in the epidermis has been suggested, no direct information regarding their nature has been available. Using cDNAs for these G-proteins as a probe, we investigated the nature of the G-proteins of various keratinocytes. FRSK cells, a cell line derived from fetal rat epidermal cells, and SV-40-transformed human epidermal cells, both of which are highly proliferative keratinocytes, contained mRNAs for both Gs-α and two Gi-α proteins (Gi-2α and -3α). No evidence for the presence of Gi-1α or Go-α was obtained. Normal human or pig epidermis had a relatively small amount of mRNAs for these G-proteins in the stable (hypoproliferative) condition. Tape-stripping-induced, and UVB-induced hyperproliferative epidermis contained an increased amount of these G-protein messages. However, all Gs-α, Gi-2α and -3α, as well as Β-actin mRNAs, were increased to a similar extent, and no specific expression of G-protein messages could be detected in the hyperproliferative epidermis.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Similar content being viewed by others

References

  1. Bray P, Carter A, Simons C, Guo V, Puckett C, Kamholz J, Spiegel A, Nirenberg M (1986) Human cDNA clones for four species of Gαs signal transduction protein. Proc Natl Acad Sci USA 83:8893–8897

    Google Scholar 

  2. Cantiello HF, Patenaude CR, Ausiello DA (1989) G protein subunit, αi-3, activates a pertussis toxin-sensitive Na+ channel from the epithelial cell line, A-6. J Biol Chem 264:20867–20870

    Google Scholar 

  3. Casey PJ, Gilman AG (1988) G protein involvement in receptor-effector coupling. J Biol Chem 263:2577–2580

    Google Scholar 

  4. Choi EJ, Toscano WA (1988) Modulation of adenylate cyclase in human keratinocytes by protein kinase C. J Biol Chem 263:17167–17172

    Google Scholar 

  5. Codina J, Yatani A, Grenet D, Brown AM, Birnbaumer L (1987) The α subunit of the GTP binding protein Gk opens atrial potassium channels. Science 236:442–445

    Google Scholar 

  6. Gilman AG (1987) G proteins: transducers of receptor-generated signals. Annu Rev Biochem 56:615–649

    Google Scholar 

  7. Iizuka H, Matsuo S, Tamura T, Ohkuma N (1988) Increased cholera toxin-, and forskolin-induced cyclic AMP accumulations in psoriatic involved versus uninvolved or normal human epidermis. J Invest Dermatol 91:154–157

    Google Scholar 

  8. Indo K, Wilson RB (1977) Fetal rat keratinizing epidermal cells in culture: effects of long term treatment by benzo[a]pyrene on their growth characteristics. J Natl Cancer Inst 59:867–880

    Google Scholar 

  9. Tsutsui M, Iizuka H (1990) Inhibitory guanine nucleotide binding protein in pig epidermis: regulation of epidermal adenylate cyclase. Arch Dermatol Res (in press)

  10. Itoh H, Katada T, Ui M, Kawasaki H, Suzuki K, Kaziro Y (1988) Identification of three pertussis toxin substrates (41, 40 and 39 kDa proteins) in mammalian brain. FEBS Lett 230:85–89

    Google Scholar 

  11. Itoh H, Toyama R, Kozasa T, Tsukamoto T, Matsuoka M, Kaziro Y (1988) Presence of three distinct molecular species of Gi protein α subunit. J Biol Chem 263:6656–6664

    Google Scholar 

  12. Jelsema CL, Axelrod J (1987) Stimulation of phospholipase A2 activity in bovine rod outer segments by the Βγ subunits of transducin and its inhibition by the α subunit. Proc Natl Acad Sci USA 84:3623–3627

    Google Scholar 

  13. Katada T, Northup JK, Bokoch GM, Ui M, Gilman AG (1984) The inhibitory guanine nucleotide-binding regulatory component of adenylate cyclase. J Biol Chem 259:3578–3585

    Google Scholar 

  14. Katada T, Oinuma M, Ui M (1986) Mechanisms for inhibition of the catalytic activity of adenylate cyclase by the guanine nucleotide-binding proteins serving as the substrate of isletactivating protein pertussis toxin. J Biol Chem 261:5215–5221

    Google Scholar 

  15. Kikuchi A, Kozawa O, Kaibuchi K, Katada T, Ui M, Takai Y (1986) Direct evidence for involvement of a guanine nucleotide-binding protein in chemotactic peptide-stimulated formation of inositol bisphosphate and trisphosphate in differentiated human leukemic (HL-60) cells. J Biol Chem 261:11 558–11 562

    Google Scholar 

  16. Kim S, Ang S-L, Bloch DB, Bloch KD, Kawahara Y, Tolman C, Lee R, Seidman JG, Neer EJ (1988) Identification of cDNA encoding an additional α subunit of a human GTP-binding protein: expression of three αi subtypes in human tissues and cell lines. Proc Natl Acad Sci USA 85:4153–4157

    Google Scholar 

  17. Kim D, Lewis DL, Graziadei L, Neer EJ, Bar-Sagi D, Clapham DE (1989) G-protein Βγ-subunits activate the cardiac muscarinic K+-channel via phospholipase A2. Nature 337:557–560

    Google Scholar 

  18. Kozasa T, Itoh H, Tsukamoto T, Kaziro Y (1988) Isolation and characterization of the human Gsα gene. Proc Natl Acad Sci USA 85:2081–2085

    Google Scholar 

  19. Lochrie MA, Simon MI (1988) G protein multiplicity in eukaryotic signal transduction systems. Biochemistry 27:4957–4965

    Google Scholar 

  20. Maniatis T, Fritsch EF, Sambrook T (1989) Molecular cloning: a laboratory manual, 2nd edn. Cold Spring Harbor Laboratory, Cold Spring Harbor, New York

    Google Scholar 

  21. Neer EJ, Clapham DE (1988) Roles of G protein subunits in transmembrane signalling. Nature 333:129–134

    Google Scholar 

  22. Ohkuma N, Kajita S, Iizuka H (1987) Superoxide dismutase in epidermis: its relation to keratinocyte proliferation. J Dermatol (Tokyo) 14:562–568

    Google Scholar 

  23. Oinuma M, Katada T, Ui M (1987) A new GTP-binding protein in differentiated human leukemic (HL-60) cells serving as the specific substrate of islet-activating protein, pertussis toxin. J Biol Chem 262:8347–8353

    Google Scholar 

  24. Robishaw JD, Smigel MD, Gilman AG (1986) Molecular basis for two froms of the G protein that stimulates adenylate cyclase. J Biol Chem 261:9587–9590

    Google Scholar 

  25. Steinberg ML, Defendi V (1983) Transformation and immortalization of human keratinocytes by SV40. J Invest Dermatol 81:131S-136S

    Google Scholar 

  26. Strathman M, Wilkie TM, Simon MI (1989) Diversity of the G-protein family: sequences from five additional alpha subunits in the mouse. Proc Natl Acad Sci USA 86:7407–7409

    Google Scholar 

  27. Thomas PS (1980) Hybridization of denatured RNA and small DNA fragments transferred to nitrocellulose. Proc Natl Acad Sci USA 77:5201–5205

    Google Scholar 

  28. Watanabe M, Iizuka H (1987) Adenylate cyclase-cyclic AMP system in pure epidermis isolated by use of dispase. J Dermatol (Tokyo) 14:336–342

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Rights and permissions

Reprints and permissions

About this article

Cite this article

Takahashi, H., Miyokawa, N., Katagiri, M. et al. Expression of guanine nucleotide binding proteins, Gs and Gi, in mRNAs in epidermal keratinocytes. Arch Dermatol Res 282, 392–396 (1990). https://doi.org/10.1007/BF00372090

Download citation

  • Received:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00372090

Key words

Navigation