Abstract
To study the process of hormone action, we have developed an in vitro system utilizing minced oviduct from estrogen-treated chicks incubated in tissue culture medium. Progesterone added to the medium induced synthesis of a specific protein, avidin, that continued for up to 96 hr. During this period there was no increase in total oviduct protein, ovalbumin, or lysozyme, which suggests the specificity of the progesterone effect. The induction process was dependent on new protein synthesis, since cycloheximide inhibited the induction completely. Actinomycin D in doses that prevented nuclear RNA synthesis, but not general protein synthesis, inhibited avidin production 70-90%. Avidin synthesis was not affected by 5-fluorouracil. The rate of DNA synthesis examined by thymidine-3H pulse labeling was not stimulated during avidin induction. Hydroxyurea (an inhibitor of DNA synthesis) and colchicine (a mitotic inhibitor) did not prevent induction. Studies utilizing uridine-3H pulses showed an effect on rapdly labeled nuclear RNA coincident with induction. Nuclear RNA polymerase activity increased before avidin induction. Since avidin was the only new protein synthesized in response to progesterone, the early stimulation of nuclear RNA synthesis and RNA polymerase activity would suggest a mechanism of action for this steroid at the transcription level of protein synthesis.
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Selected References
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- BURTON K. A study of the conditions and mechanism of the diphenylamine reaction for the colorimetric estimation of deoxyribonucleic acid. Biochem J. 1956 Feb;62(2):315–323. doi: 10.1042/bj0620315. [DOI] [PMC free article] [PubMed] [Google Scholar]
- ENNIS H. L., LUBIN M. CYCLOHEXIMIDE: ASPECTS OF INHIBITION OF PROTEIN SYNTHESIS IN MAMMALIAN CELLS. Science. 1964 Dec 11;146(3650):1474–1476. doi: 10.1126/science.146.3650.1474. [DOI] [PubMed] [Google Scholar]
- FRENKEL E. P., SKINNER W. N., SMILEY J. D. STUDIES ON A METABOLIC DEFECT INDUCED BY HYDROXYUREA (NSC-32065). Cancer Chemother Rep. 1964 Aug;40:19–22. [PubMed] [Google Scholar]
- GARREN L. D., HOWELL R. R., TOMKINS G. M., CROCCO R. M. A PARADOXICAL EFFECT OF ACTINOMYCIN D: THE MECHANISM OF REGULATION OF ENZYME SYNTHESIS BY HYDROCORTISONE. Proc Natl Acad Sci U S A. 1964 Oct;52:1121–1129. doi: 10.1073/pnas.52.4.1121. [DOI] [PMC free article] [PubMed] [Google Scholar]
- GARREN L. D., HOWELL R. R., TOMKINS G. M. MAMMALIAN ENZYME INDUCTION BY HYDROCORTISONE: THE POSSIBLE ROLE OF RNA. J Mol Biol. 1964 Jul;9:100–108. doi: 10.1016/s0022-2836(64)80093-0. [DOI] [PubMed] [Google Scholar]
- GORSKI J. EARLY ESTROGEN EFFECTS ON THE ACTIVITY OF UTERINE RIBONUCLEIC ACID POLYMERASE. J Biol Chem. 1964 Mar;239:889–892. [PubMed] [Google Scholar]
- HOROWITZ J., SAUKKONEN J. J., CHARGAFF E. Effect of 5-fluorouracil on a uracil-requiring mutant of Escherichia coli. Biochim Biophys Acta. 1958 Jul;29(1):222–223. doi: 10.1016/0006-3002(58)90175-6. [DOI] [PubMed] [Google Scholar]
- Henshaw E. C., Revel M., Hiatt H. H. A cytoplasmic particle bearing messenger ribonucleic acid in rat liver. J Mol Biol. 1965 Nov;14(1):241–256. doi: 10.1016/s0022-2836(65)80244-3. [DOI] [PubMed] [Google Scholar]
- Ilan J., Quastel J. H. Effects of colchicine on nucleic acid metabolism during metamorphosis of Tenebrio molitor L. and in some mammalian tissues. Biochem J. 1966 Aug;100(2):448–457. doi: 10.1042/bj1000448. [DOI] [PMC free article] [PubMed] [Google Scholar]
- JACOB F., MONOD J. Genetic regulatory mechanisms in the synthesis of proteins. J Mol Biol. 1961 Jun;3:318–356. doi: 10.1016/s0022-2836(61)80072-7. [DOI] [PubMed] [Google Scholar]
- KORN D., PROTASS J. J., LEIVE L. A NOVEL EFFECT OF ACTINOMYCIN D IN PREVENTING BACTERIOPHAGE T4 MATURATION IN ESCHERICHIA COLI. Biochem Biophys Res Commun. 1965 May 3;19:473–481. doi: 10.1016/0006-291x(65)90149-x. [DOI] [PubMed] [Google Scholar]
- LANG N., SEKERKIS C. E. STIMULATION OF RNA-POLYMERASE ACTIVITY IN RAT LIVER BY CORTISOL. Life Sci. 1964 May;3:391–393. doi: 10.1016/0024-3205(64)90198-5. [DOI] [PubMed] [Google Scholar]
- LITTLEFIELD J. W., KELLER E. B., GROSS J., ZAMECNIK P. C. Studies on cytoplasmic ribonucleoprotein particles from the liver of the rat. J Biol Chem. 1955 Nov;217(1):111–123. [PubMed] [Google Scholar]
- LITWACK G. Photometric determination of lysozyme activity. Proc Soc Exp Biol Med. 1955 Jul;89(3):401–403. doi: 10.3181/00379727-89-21824. [DOI] [PubMed] [Google Scholar]
- LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
- Laszlo J., Miller D. S., McCarty K. S., Hochstein P. Actinomycin D: inhibition of respiration and glycolysis. Science. 1966 Feb 25;151(3713):1007–1010. doi: 10.1126/science.151.3713.1007. [DOI] [PubMed] [Google Scholar]
- Liao S., Leininger K. R., Sagher D., Barton R. W. Rapid effect of testosterone on ribonucleic acid polymerase activity of rat ventral prostate. Endocrinology. 1965 Oct;77(4):763–765. doi: 10.1210/endo-77-4-763. [DOI] [PubMed] [Google Scholar]
- O'Malley B. W. In vitro hormonal induction of a specific protein (avidin) in chick oviduct. Biochemistry. 1967 Aug;6(8):2546–2551. doi: 10.1021/bi00860a036. [DOI] [PubMed] [Google Scholar]
- O'Malley B. W., McGuire W. L., Middleton P. A. Structure-function relationships of various steroids relative to induction of a specific oviduct protein (Avidin). Endocrinology. 1967 Sep;81(3):677–678. doi: 10.1210/endo-81-3-677. [DOI] [PubMed] [Google Scholar]
- REVEL M., HIATT H. H., REVEL J. P. ACTINOMYCIN D: AN EFFECT ON RAT LIVER HOMOGENATES UNRELATED TO ITS ACTION ON RNA SYNTHESIS. Science. 1964 Dec 4;146(3649):1311–1313. doi: 10.1126/science.146.3649.1311. [DOI] [PubMed] [Google Scholar]
- Roberts W. K., Newman J. F. Use of low concentrations of actinomycin D in the study of RNA synthesis in Ehrlich ascites cells. J Mol Biol. 1966 Sep;20(1):63–73. doi: 10.1016/0022-2836(66)90117-3. [DOI] [PubMed] [Google Scholar]
- Shearer R. W., McCarthy B. J. Evidence for ribonucleic acid molecules restricted to the cell nucleus. Biochemistry. 1967 Jan;6(1):283–289. doi: 10.1021/bi00853a044. [DOI] [PubMed] [Google Scholar]
- Stockdale F. E., Topper Y. J. The role of DNA synthesis and mitosis in hormone-dependent differentiation. Proc Natl Acad Sci U S A. 1966 Oct;56(4):1283–1289. doi: 10.1073/pnas.56.4.1283. [DOI] [PMC free article] [PubMed] [Google Scholar]
- TOMKINS G. M., MAXWELL E. S. SOME ASPECTS OF STEROID HORMONE ACTION. Annu Rev Biochem. 1963;32:677–708. doi: 10.1146/annurev.bi.32.070163.003333. [DOI] [PubMed] [Google Scholar]
- Tata J. R. Hormones and the synthesis and utilization of ribonucleic acids. Prog Nucleic Acid Res Mol Biol. 1966;5:191–250. doi: 10.1016/s0079-6603(08)60235-4. [DOI] [PubMed] [Google Scholar]
- Tomkins G. M., Thompson E. B., Hayashi S., Gelehrter T., Granner D., Peterkofsky B. Tyrosine transaminase induction in mammalian cells in tissue culture. Cold Spring Harb Symp Quant Biol. 1966;31:349–360. doi: 10.1101/sqb.1966.031.01.045. [DOI] [PubMed] [Google Scholar]
- WEBER G., SRIVASTAVA S. K., SINGHAL R. L. ROLE OF ENZYMES IN HOMEOSTASIS. VII. EARLY EFFECTS OF CORTICOSTEROID HORMONES ON HEPATIC GLUCONEOGENIC ENZYMES, RIBONUCLEIC ACID METABOLISM, AND AMINO ACID LEVEL. J Biol Chem. 1965 Feb;240:750–756. [PubMed] [Google Scholar]
- WILLIAMS-ASHMAN H. G., LIAO S., HANCOCK R. L., JURKOWITZ L., SILVERMAN D. A. TESTICULAR HORMONES AND THE SYNTHESIS OF RIBONUCLEIC ACIDS AND PROTEINS IN THE PROSTATE GLAND. Recent Prog Horm Res. 1964;20:247–301. [PubMed] [Google Scholar]
- Widnell C. C., Tata J. R. Additive effects of thyroid hormone, growth hormone and testosterone on deoxyribonucleic acid-dependent ribonucleic acid polymerase in rat-liver nuclei. Biochem J. 1966 Feb;98(2):621–629. doi: 10.1042/bj0980621. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Williams-Ashman H. G. New facets of the biochemistry of steroid hormone action. Cancer Res. 1965 Aug;25(7):1096–1124. [PubMed] [Google Scholar]
- YOUNG C. W., HODAS S. HYDROXYUREA: INHIBITORY EFFECT ON DNA METABOLISM. Science. 1964 Nov 27;146(3648):1172–1174. doi: 10.1126/science.146.3648.1172. [DOI] [PubMed] [Google Scholar]