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.
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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
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
Casey PJ, Gilman AG (1988) G protein involvement in receptor-effector coupling. J Biol Chem 263:2577–2580
Choi EJ, Toscano WA (1988) Modulation of adenylate cyclase in human keratinocytes by protein kinase C. J Biol Chem 263:17167–17172
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
Gilman AG (1987) G proteins: transducers of receptor-generated signals. Annu Rev Biochem 56:615–649
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
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
Tsutsui M, Iizuka H (1990) Inhibitory guanine nucleotide binding protein in pig epidermis: regulation of epidermal adenylate cyclase. Arch Dermatol Res (in press)
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
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
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
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
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
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
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
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
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
Lochrie MA, Simon MI (1988) G protein multiplicity in eukaryotic signal transduction systems. Biochemistry 27:4957–4965
Maniatis T, Fritsch EF, Sambrook T (1989) Molecular cloning: a laboratory manual, 2nd edn. Cold Spring Harbor Laboratory, Cold Spring Harbor, New York
Neer EJ, Clapham DE (1988) Roles of G protein subunits in transmembrane signalling. Nature 333:129–134
Ohkuma N, Kajita S, Iizuka H (1987) Superoxide dismutase in epidermis: its relation to keratinocyte proliferation. J Dermatol (Tokyo) 14:562–568
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
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
Steinberg ML, Defendi V (1983) Transformation and immortalization of human keratinocytes by SV40. J Invest Dermatol 81:131S-136S
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
Thomas PS (1980) Hybridization of denatured RNA and small DNA fragments transferred to nitrocellulose. Proc Natl Acad Sci USA 77:5201–5205
Watanabe M, Iizuka H (1987) Adenylate cyclase-cyclic AMP system in pure epidermis isolated by use of dispase. J Dermatol (Tokyo) 14:336–342
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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
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DOI: https://doi.org/10.1007/BF00372090