[en] Expression of the rat growth hormone (rGH) gene in the pituitary and in cultured pituitary tumor cells is regulated by glucocorticoid hormones. After co-transfer of cloned DNA containing the rGH gene with the herpes simplex virus (HSV) thymidine kinase (tk) gene into mouse Ltk- cells, rGH gene transcripts were detected in eight of fifteen tk+ cell lines. However, in all eight clones, the predominant rGH gene transcript was only about 0.75 kb, 0.3 kb shorter than pituitary rGH mRNA. The 0.75-kb transcripts, examined from one clone, L-rGH-4, lacked sequences derived from exons 1 and 2 of the rGH gene. Although transcripts larger than 0.75 kb were detected, the normal 2.2-kb rGH gene primary transcript was present only at very low levels. Nuclease mapping studies also failed to reveal transcripts initiated at the normal rGH gene promoter, but instead revealed transcripts with 5' termini arising within intron B of the gene. These data suggest either that transcripts arise from internal promoters within the rGH gene or that a transcript initiated upstream from the normal promoter was processed abnormally. Dexamethasone increased the levels of the 0.75-kb rGH gene transcripts about fourfold in all eight clones expressing rGH mRNA. These data suggest that structural elements important for glucocorticoid-mediated influences on regulation of GH gene expression are contained within the transferred rGH gene fragment and can function even when the normal rGH gene promoter is not used and the pattern of expression is grossly abnormal.
Disciplines :
Biochemistry, biophysics & molecular biology
Author, co-author :
Karin, M.
Eberhardt, N. L.
Mellon, S. H.
Malich, N.
Richards, R. I.
Slater, E. P.
Barta, A.
Martial, Joseph ; Université de Liège - ULiège > Département des sciences de la vie > GIGA-R : Biologie et génétique moléculaire
Baxter, J. D.
Cathala, G.
Language :
English
Title :
Expression and hormonal regulation of the rat growth hormone gene in transfected mouse L cells
Publication date :
1984
Journal title :
DNA
ISSN :
0198-0238
Publisher :
Mary Ann Liebert, Inc., New York, United States - New York
ANDERSON, C.W., LEWIS, J.B., ATKINS, J.F., and GESTELAND, R.F. (1974). Cell-free synthesis of adenovirus 2 proteins programmed by fractionated messenger RNA: A comparison of polypeptide products and messenger RNA lengths. Proc. Natl. Acad. Sci. USA 71, 2756–2760.
AVIV, H., and LEDER, P. (1972). Purification of biologically active globin messenger RNA by chromatography on oligothymidylic acid-cellulose. Proc. Natl. Acad. Sci. USA 69, 1408–1412.
BANCROFT, F.C., LEVINE, L., and TASHJIAN, A.H., Jr. (1969). Control of growth hormone production by a clonal strain of rat pituitary cells. Stimulation by hydrocortisone. J. Cell Biol. 43, 432–441.
BARTA, A., RICHARDS, R.I., BAXTER, J.D., and SHINE, J. (1981). Primary structure and evolution of rat growth hormone gene. Proc. Natl. Acad. Sci. USA 78, 4867–4871.
BREATHNACH, R., MANTEI, N., and CHAMBON, P. (1980). Correct splicing of a chicken ovalbumin gene transcript in mouse L cells. Proc. Natl. Acad. Sci. USA 77, 740–744.
BROCK, M.L., and SHAPIRO, D.J. (1983). Estrogen stabilizers vitellogenin mRNA against cytoplasmic degradation. Cell 34, 207–214.
BUETTI, E., and DIGGELMANN, H. (1981). Cloned mouse mammary tumor virus DNA is biologically active in transfected mouse cells and its expression is stimulated by glucocorticoid hormones. Cell 23, 335–345.
CATHALA, G., SAVOURET, J.-F., MENDEZ, B., WEST, B.L., KARIN, M., MARTIAL, J.A., and BAXTER, J.D. (1983). A method for isolation of intact and translationally active ribonucleic acid. DNA 2, 327–333.
CHIRGWIN, J.M., PRZYBYLA, A.E., MacDONALD, R.J., and RUTTER, W.J. (1979). Isolation of biologically active ribonucleic acid from sources enriched in ribonuclease. Biochemistry 18, 5294–5299.
COX, R.F. (1977). Estrogen withdrawal in chick oviduct. Selective loss of high abundance classes of polyadenylated messenger RNA. Biochemistry 16, 3433–3443.
DOBNER, P.R., KAWASAKI, E.S., YU, L.-Y., and BANCROFT, F.C. (1981). Thyroid or glucocorticoid hormone induces pre-growth-hormone mRNA and its probable nuclear precursor in rat pituitary cells. Proc. Natl. Acad. Sci. USA 78, 2230–2234.
DOEHMER, J., BARINAGA, M., VALE, W., ROSENFELD, M.G., VERMA, I.M., and EVANS, R.M. (1982). Introduction of rat growth hormone gene into mouse fibroblasts via a retroviral DNA vector: Expression and regulation. Proc. Natl. Acad. Sci. USA 79, 2268–2272.
EBERHARDT, N.L., APRILETTI, J.A., and BAXTER, J.D. (1980). Molecular biology of thyroid hormone action. In G. Litwack, Ed. The Biochemical Actions of Hormones, Vol. 7 (Academic Press, New York) pp. 311–393.
EVANS, R.M., BIRNBERG, N.C., and ROSENFELD, M.G. (1982). Glucocorticoid and thyroid hormones transcriptionally regulate growth hormone gene expression. Proc. Natl. Acad. Sci. USA 79, 7659–7663.
GRAHAM, F.L., BACCHETTI, S., McKINNON, R., STANNERS, C., CORDELL, B., and GOODMAN, H.M. (1980). Transformation of mammalian cells with DNA using the calcium technique. In R. Baserga, C. Croce, and G. Rovera, Eds. Introduction of Macromolecules into Viable Mammalian Cells (Alan R. Liss, New York) pp. 3–25.
HERLANT, M. (1964). The cells of the adenohypophysis and their functional significance. Int. Rev. Cytol. 17, 299–382.
HUANG, A.L., OSTROWSKI, M.C., BERARD, D., and HAGER, G.L. (1981). Glucocorticoid regulation of the Ha-MuSV p21 gene conferred by sequences from mouse mammary tumor virus. Cell 27, 245–255.
HYNES, N.E., KENNEDY, N., RAHMSDORF, U., and GRONER, B. (1981). Homome-responsive expression of an endogenous proviral gene of mouse mammary tumor virus after molecular cloning and gene transfer into cultured cells. Proc. Natl. Acad. Sci. USA 78, 2038–2042.
KARIN, M., ANDERSEN, R.D., and HERSCHMAN, H.R. (1981). Induction of metallothionein mRNA in HeLa cells by dexamethasone and by heavy metals. Eur. J. Biochem. 118, 527–531.
KARIN, M., HASLINGER, A., HOLTGREVE, H., CATHALA, G., SLATER, E., and BAXTER, J.D. (1984). Activation of a heterologous promoter in response to dexamethasone and cadmium by metallothionein gene 5′-flanking DNA. Cell (in press).
KARIN, M., and RICHARDS, R.I. (1982). Human metallothionein genes-primary structure of the metallothionein-II gene and a related processed gene. Nature 299, 797–802.
KURTZ, D.T. (1981). Hormonal inducibility of rat α2u globulin genes in transfected mouse cells. Nature 291, 629–631.
LAI, E.C., WOO, S.L.C., BORDELON-RISER, M.E., FRASER, T.H., and O'MALLEY, B.W. (1980). Ovalbumin is synthesized in mouse cells transformed with the natural chicken ovalbumin gene. Proc. Natl. Acad. Sci. USA 77, 244–248.
LEE, F., MULLIGAN, R., BERG, P., and RINGOLD, G. (1981). Glucocorticoids regulate expression of dihydrofolate reductase cDNA in mouse mammary tumor virus chimaeric plasmids. Nature 294, 228–232.
LITTLEFIELD, J.W. (1963). The inosinic acid pyrophosphory-lase activity of mouse fibroblasts partially resistant to 8-azaguanine. Proc. Natl. Acad. Sci. USA 50, 568–576.
MANTEI, N., BOLL, W., and WEISSMANN, C. (1979). Rabbit β-globin mRNA production in mouse L cells transformed with cloned rabbit β-globin chromosomal DNA. Nature 281, 40–46.
MARTIAL, J.A., BAXTER, J.D., GOODMAN, H.M., and SEEBURG, P.H. (1977a). Regulation of growth hormone messenger RNA by thyroid and glucocorticoid hormones. Proc. Natl. Acad. Sci. USA 74, 1816–1820.
MARTIAL, J.A., SEEBURG, P.H., GUENZI, D., GOODMAN, H.M., and BAXTER, J.D. (1977b). Regulation of growth hormone gene expression: Synergistic effects of thyroid and glucocorticoid hormones. Proc. Natl. Acad. Sci. USA 74, 4293–4295.
McKNIGHT, G.S., and PALMITER, R.D. (1979). Transcriptional regulation of the ovalbrimin and conallbrimin genes by steroid hormones in chick oviduct. J. Biol. Chem. 254, 9050–9058.
O'FARRELL, P.H., KUTTER, E., and NAKANISHI, M. (1980). A restriction map of the bacteriophage T4 genome. Molec. Gen. Genet. 179, 421–435.
RENKAWITZ, R., BEUG, H., GRAF, T., MATTHIAS, P., GREZ, M., and SCHUTZ, G. (1982). Expression of a chicken lysozyme recombinant gene is regulated by progesterone and dexamethasone after microinjection into oviduct cells. Cell 31, 167–176.
ROBINS, D.M., PAEK, I., SEEBURG, P.H., and AXEL, R. (1982). Regulated expression of human growth hormone genes in mouse cells. Cell 29, 623–631.
SEEBURG, P.H., SHINE, J., MARTIAL, J.A., BAXTER, J.D., and GOODMAN, H.M. (1977). Nucleotide sequence and amplification in bacteria of structural gene for rat growth hormone. Nature 270, 486–494.
SHAPIRO, L.E., SAMUELS, H.H., and YAFFE, B.M. (1978). Thyroid and glucocorticoid hormones synergistically control growth hormone mRNA in cultured GHi cells. Proc. Natl. Acad. Sci. 75, 45–49.
SPINDLER, S.R., MELLON, S.H., and BAXTER, J.D. (1982). Growth hormone gene transcription is regulated by thyroid and glucocorticoid hormones in cultured rat pituitary tumor cells. J. Biol. Chem. 257, 11627–11632.
THOMAS, P.S. (1980). Hybridization of denatured RNA and small DNA fragments transferred to nitrocellulose. Proc. Natl. Acad. Sci. USA 77, 5201–5205.
UCKER, D.S., ROSS, S.R., and YAMAMOTO, K.R. (1981). Mammary tumor virus DNA contains sequences required for its hormone-regulated transcription. Cell 27, 257–266.
WEAVER, R.F., and WEISSMANN, C. (1979). Mapping of RNA by a modification of the Berk-Sharp procedure: The 5′ termini of 15 S β-globin mRNA precursor and mature 10 S β-globin mRNA have identical map coordinates. Nucleic Acids Res. 7, 1175–1193.
WEGNEZ, M., SCHACHTER, B.S., BAXTER, J.D., and MARTIAL, J.A. (1982). Hormonal regulation of growth hormone mRNA. DNA 1, 145–153.
WIGLER, M., SWEET, R., SIM, G.K., WOLD, B., PELLICER, A., LACY, E., MANIATIS, T., SILVERSTEIN, S., and AXEL, R. (1979). Transformation of mammalian cells with genes from procaryotes and eucaryotes. Cell 16, 777–785.
WOLD, B., WIGLER, M., LACY, E., MANIATIS, T., SILVERSTEIN, S., and AXEL, R. (1979). Introduction and expression of a rabbit β-globin gene in mouse fibroblasts. Proc. Natl. Acad. Sci. USA 76, 5684–5688.
YASUMURA, Y., TASHJIAN, A.J. Jr., and SATO, G.H. (1966). Establishment of four functional, clonal strains of animal cells in culture. Science 154, 1186–1189.