Skip to main content
PMC 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
. 1971 Aug;68(8):1725–1729. doi: 10.1073/pnas.68.8.1725

Gene Activation in WI-38 Fibroblasts Stimulated to Proliferate: Requirement for Protein Synthesis

Giovanni Rovera 1, John Farber 1, Renato Baserga 1
PMCID: PMC389280  PMID: 5288757

Abstract

Confluent monolayers of WI-38 human diploid fibroblasts can be stimulated to divide by fresh medium containing 30% fetal-calf serum. Up to 80% of the cells are stimulated to divide, with a peak of DNA synthesis between 15 and 21 hr. 1 hr after the change of medium there is a 70% rise in chromatin template activity. Cycloheximide inhibited the increase in chromatin template activity. A requirement for RNA synthesis was investigated by incubating stimulated and unstimulated cells with 10 μg/ml of actinomycin D. In spite of a 95% inhibition of RNA synthesis in whole cells, purified chromatin from stimulated cells showed the usual increase in template activity. These experiments implicate a requirement for protein synthesis in template activation, and imply that the synthesis of this (or these) protein(s) is independent of RNA synthesis and regulated by a purely translational mechanism.

Keywords: template activation, human diploid fibroblasts, cell proliferation

Full text

PDF

Selected References

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

  1. ALLFREY V. G., FAULKNER R., MIRSKY A. E. ACETYLATION AND METHYLATION OF HISTONES AND THEIR POSSIBLE ROLE IN THE REGULATION OF RNA SYNTHESIS. Proc Natl Acad Sci U S A. 1964 May;51:786–794. doi: 10.1073/pnas.51.5.786. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. ALLFREY V. G., LITTAU V. C., MIRSKY A. E. On the role of of histones in regulation ribonucleic acid synthesis in the cell nucleus. Proc Natl Acad Sci U S A. 1963 Mar 15;49:414–421. doi: 10.1073/pnas.49.3.414. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. 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]
  4. Baserga R., Estensen R. D., Petersen R. O. Inhibition of DNA synthesis in Ehrlich ascites cells by actinomycin D. II. The presynthetic block in the cell cycle. Proc Natl Acad Sci U S A. 1965 Oct;54(4):1141–1148. doi: 10.1073/pnas.54.4.1141. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Bekhor I., Kung G. M., Bonner J. Sequence-specific interaction of DNA and chromosomal protein. J Mol Biol. 1969 Jan;39(2):351–364. doi: 10.1016/0022-2836(69)90322-2. [DOI] [PubMed] [Google Scholar]
  6. CHAMBERLIN M., BERG P. Deoxyribo ucleic acid-directed synthesis of ribonucleic acid by an enzyme from Escherichia coli. Proc Natl Acad Sci U S A. 1962 Jan 15;48:81–94. doi: 10.1073/pnas.48.1.81. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Chaudhuri S., Lieberman I. Control of ribosome syntesis in normal and regenerating liver. J Biol Chem. 1968 Jan 10;243(1):29–33. [PubMed] [Google Scholar]
  8. Farber J., Rovera G., Baserga R. Template activity of chromatin during stimulation of cellular proliferation in human diploid fibroblasts. Biochem J. 1971 Apr;122(2):189–195. doi: 10.1042/bj1220189. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Gilman A. G. A protein binding assay for adenosine 3':5'-cyclic monophosphate. Proc Natl Acad Sci U S A. 1970 Sep;67(1):305–312. doi: 10.1073/pnas.67.1.305. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. HUANG R. C., BONNER J. Histone, a suppressor of chromosomal RNA synthesis. Proc Natl Acad Sci U S A. 1962 Jul 15;48:1216–1222. doi: 10.1073/pnas.48.7.1216. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Hearst J. E., Botchan M. The eukaryotic chromosome. Annu Rev Biochem. 1970;39:151–182. doi: 10.1146/annurev.bi.39.070170.001055. [DOI] [PubMed] [Google Scholar]
  12. Kleinsmith L. J., Allfrey V. G., Mirsky A. E. Phosphoprotein metabolism in isolated lymphocyte nuclei. Proc Natl Acad Sci U S A. 1966 May;55(5):1182–1189. doi: 10.1073/pnas.55.5.1182. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. LEVINE E. M., BECKER Y., BOONE C. W., EAGLE H. CONTACT INHIBITION, MACROMOLECULAR SYNTHESIS, AND POLYRIBOSOMES IN CULTURED HUMAN DIPLOID FIBROBLASTS. Proc Natl Acad Sci U S A. 1965 Feb;53:350–356. doi: 10.1073/pnas.53.2.350. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. LIEBERMAN I., ABRAMS R., OVE P. Changes in the metabolism of ribonucleic acid preceding the synthesis of deoxyribonucleic acid in mammalian cells cultured from the animal. J Biol Chem. 1963 Jun;238:2141–2149. [PubMed] [Google Scholar]
  15. 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]
  16. Neal G. E., Ali-Khan M. Y., Mangan F. R. The effect of cycloheximide on RNA synthesis in regenerating rat liver. Biochim Biophys Acta. 1970 May 21;209(1):257–259. doi: 10.1016/0005-2787(70)90683-0. [DOI] [PubMed] [Google Scholar]
  17. PATTERSON M. S., GREENE R. C. MEASUREMENT OF LOW ENERGY BETA-EMITTERS IN AQUEOUS SOLUTION BY LIQUID SCINTILLATION COUNTING OF EMULSIONS. Anal Chem. 1965 Jun;37:854–857. doi: 10.1021/ac60226a017. [DOI] [PubMed] [Google Scholar]
  18. Paul J., Gilmour R. S. Organ-specific restriction of transcription in mammalian chromatin. J Mol Biol. 1968 Jul 14;34(2):305–316. doi: 10.1016/0022-2836(68)90255-6. [DOI] [PubMed] [Google Scholar]
  19. Pogo A. O., Allfrey V. G., Mirsky A. E. Evidence for increased DNA template activity in regenerating liver nuclei. Proc Natl Acad Sci U S A. 1966 Aug;56(2):550–557. doi: 10.1073/pnas.56.2.550. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Rhode S. L., 3rd, Ellem K. A. Control of nucleic acid synthesis in human diploid cells undergoing contact inhibition. Exp Cell Res. 1968 Oct;53(1):184–204. doi: 10.1016/0014-4827(68)90366-2. [DOI] [PubMed] [Google Scholar]
  21. Rovera G., Baserga R. Early changes in the synthesis of acidic nuclear proteins in human diploid fibroblasts stimulated to synthesize DNA by changing the medium. J Cell Physiol. 1971 Apr;77(2):201–211. doi: 10.1002/jcp.1040770211. [DOI] [PubMed] [Google Scholar]
  22. SCOTT J. F., FRACCASTORO A. P., TAFT E. B. Studies in histochemistry. I. Determination of nucleic acids in microgram amounts of tissue. J Histochem Cytochem. 1956 Jan;4(1):1–10. doi: 10.1177/4.1.1. [DOI] [PubMed] [Google Scholar]
  23. Stein G., Baserga R. Continued synthesis of non-histone chromosomal proteins during mitosis. Biochem Biophys Res Commun. 1970 Nov 9;41(3):715–722. doi: 10.1016/0006-291x(70)90072-0. [DOI] [PubMed] [Google Scholar]
  24. Stein G., Baserga R. The synthesis of acidic nuclear proteins in the prereplicative phase of the isoproterenol-stimulated salivary gland. J Biol Chem. 1970 Nov 25;245(22):6097–6105. [PubMed] [Google Scholar]
  25. Teng C. S., Hamilton T. H. Role of chromatin in estrogen action in the uterus. II. Hormone-induced synthesis of nonhistone acidic proteins which restore histone-inhibited DNA-dependent RNA synthesis. Proc Natl Acad Sci U S A. 1969 Jun;63(2):465–472. doi: 10.1073/pnas.63.2.465. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Teng C. S., Hamilton T. H. The role of chromatin in estrogen action in the uterus, I. The control of template capacity and chemical composition and the binding of H3-estradiol-17 beta. Proc Natl Acad Sci U S A. 1968 Aug;60(4):1410–1417. doi: 10.1073/pnas.60.4.1410. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Wiebel F., Baserga R. Early alterations in amino acid pools and protein synthesis of diploid fibroblasts stimulated to synthesize DNA by addition of serum. J Cell Physiol. 1969 Oct;74(2):191–202. doi: 10.1002/jcp.1040740211. [DOI] [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