Virchows Archiv B

, Volume 17, Issue 1, pp 319–336 | Cite as

Synthesis of poly(A)-associated RNA in the prereplicative phase of isoproterenol-stimulated DNA synthesis

  • A. M. Novi


Rapidly labelled RNA containing poly(A) sequences has been isolated from polysomes of mouse parotid glands. Filtration on Millipore and binding to poly(U) immobilized on Sepharose-4B, together with significant RNase-resistance (11–16%) indicated that this polysomal fraction contains large tracts of polyadenylate sequences. A single injection of isoproterenol, a procedure that causes, after a lag period of 20 hrs, a marked stimulation of DNA synthesis and cell division in salivary glands of rodents, induces a biphasic increase in the incorporation of (3H)-adenosine into poly(A)-associated polysomal RNA. The incorporation of (3H)-adenosine is already increased within 1 hour after stimulation, decreases at 3hrs and increases again at 8 hrs, paralleling previously reported changes on chromatin template activity of mouse parotids. However, the percent poly(A) content, and the size of the poly(A)-associated RNA remain constant. Actinomycin D given 30 min prior to isoproterenol, inhibits isoproterenol-stimulated poly(A)-rich RNA synthesis. These results demonstrate that, as part of the early molecular events of its action, isoproterenol regulates transcription of adenylate-rich RNA in mouse parotid glands.

Key words

Cell Proliferation Isoproterenol Parotid Glands 


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  1. Adesnik, M., Salditt, M., Thomas, W., Darnell, J. E.: Evidence that all messenger RNA molecules (except histone messenger RNA) contain poly(A) sequences and that the poly(A) has a nuclear function. J. molec. Biol.71, 21–30 (1972)PubMedCrossRefGoogle Scholar
  2. Allfrey, V. G.: The role of chromosomal proteins in gene activation. In: Biochemistry of cell division. Baserga, R., editor, p. 179–205. Springfield, Illinois: Charles C. Thomas Publisher 1969Google Scholar
  3. Bannai, S., Terayama, H.: Template activity of chromatins isolated from regenerating rat liver. J. Biochem. (Tokyo)66, 289–295 (1969)Google Scholar
  4. Barker, K. L., Warren, J. C.: Template capacity of uterine chromatin: Control by estradiol. Proc. nat. Acad. Sci. (Wash.)56, 1298–1302 (1966)CrossRefGoogle Scholar
  5. Baserga, R., Stein, G.: Nuclear acidic proteins and cell proliferation. Fed. Proc.30, 1752–1759 (1971)PubMedGoogle Scholar
  6. Bauduin, H., Colin, M., Dumont, J. E.: Energy sources for protein synthesis and enzymatic secretion in rat pancreas in vitro. Biochim. biophys. Acta (Amst.)174, 722–733 (1969)Google Scholar
  7. Brawerman, G.: The isolation of RNA from mammalian cells. In: Methods in cell biology. Prescott, D. M., editor. New York-London: Academic Press 1973Google Scholar
  8. Brawerman G., Mendecki, J., Lee, S. Y.: A procedure for the isolation of mammalian messenger ribonucleic acid. Biochemistry11, 637–641 (1972)PubMedCrossRefGoogle Scholar
  9. Chaudhuri, S., Lieberman, I.: Control of ribosome synthesis in normal and regenerating liver. J. biol. Chem.243, 29–33 (1968)PubMedGoogle Scholar
  10. Church, R. B., McCarthy, B. J.: Unstable nuclear RNA synthesis following estrogen stimulation. Biochim. biophys. Acta (Amst.)199, 103–114 (1970)Google Scholar
  11. Couch, R. M., Anderson, K. M.: Rat ventral prostate chromatin. Effect of androgens on its chemical composition, physical properties, and template activity. Biochemistry12, 3114–3121 (1973)PubMedCrossRefGoogle Scholar
  12. Darnell, J. E., Jelinek, W. R., Molloy, G. R.: Biogenesis of mRNA: Genetic regulation in mammalian cells. Science181, 1215–1221 (1973)PubMedCrossRefGoogle Scholar
  13. Edström, J. E., Tanguay, R.: Cytoplasmic ribonucleic acids with messenger characteristics in salivary gland cells of Chironomus tentans. J. molec. Biol.84, 569–583 (1974)PubMedCrossRefGoogle Scholar
  14. Greenberg, J. R., Perry, R. P.: The isolation and characterization of steady-state labeled messenger RNA from L-cells. Biochim. biophys. Acta (Amst.)287, 361–366 (1972)Google Scholar
  15. Lee, S. Y., Mendecki, J., Brawerman, G.: A polynucleotide segment rich in adenylic acid in the rapidly-labeled polyribosomal RNA component of mouse sarcoma 180 ascites cell. Proc. nat. Acad. Sci. (Wash.)68, 1331–1335 (1971)CrossRefGoogle Scholar
  16. Lieberman, I.: On the control of mammalian deoxyribonucleic acid synthesis. In Vitro6, 46–54 (1970)PubMedCrossRefGoogle Scholar
  17. Linberg, U., Persson, T.: Isolation of mRNA from KB-cells by affinity chromatography on polyuridylic acid covalently linked to sepharose. Europ. J. Biochem.31, 246–254 (1972)CrossRefGoogle Scholar
  18. Mayfield, J. E., Bonner, J.: A partial sequence of nuclear events in regenerating rat liver. Proc. nat. Acad. Sci. (Wash.)69, 7–10 (1972)CrossRefGoogle Scholar
  19. Nakazato, H., Edmonds, M.: The isolation and purification of rapidly labeled polysome-bound ribonucleic acid on polythymidylate cellulose. J. biol. Chem.247, 3365–3367 (1972)PubMedGoogle Scholar
  20. Novi, A. M., Baserga, R.: Changes in chromatin template activity and their relationship to DNA synthesis in mouse parotid glands stimulated by isoproterenol. J. Cell Biol.55, 554–562 (1972a)PubMedCrossRefGoogle Scholar
  21. Novi, A. M., Baserga, R.: Correlation between synthesis of ribosomal RNA and stimulation of DNA synthesis in mouse salivary glands. Lab. Invest.26, 540–547 (1972b)PubMedGoogle Scholar
  22. Novi, A. M., Baserga, R.: Changes in cytoplasmic poly(A)-rich RNA in WI-38 cells stimulated to proliferate. In: 20 Anniversary Symposium on fundamental aspects of neoplasia. Rutgers University, State University of New Jersey. May 20–22. Gottlieb A. A. Editor. 1974Google Scholar
  23. Pegoraro, L., Baserga, R.: Time of appearance of deoxythymidylate kinase and deoxythymidylate synthetase and of their templates in isoproterenol-stimulated deoxyribonucleic acid synthesis. Lab. Inv.22, 226–271 (1970)Google Scholar
  24. Penman, S., Vesco, C., Penman, M.: Localization and kinetics of formation of nuclear heterodisperse RNA, cytoplasmic heterodisperse RNA and polyribosome-associated messenger RNA in HeLa cells. J. molec. Biol.34, 49–69 (1968PubMedCrossRefGoogle Scholar
  25. Perry, R. P., Kelley, D. E.: Persistent synthesis of 5S RNA when production of 28S and 18S ribosomal RNA is inhibited by low doses of Actinomycin D. J. cell. Physiol.72, 235–246 (1968)PubMedCrossRefGoogle Scholar
  26. Perry, R. P., La Torre, J., Kelley, D. E., Greenberg, G. R.: On the lability of poly(A) sequences during extraction of messenger RNA from polyribosomes. Biochim. biophys. Acta (Amst.)262, 220–226 (1972)Google Scholar
  27. Petermann, M. L.: The physical and chemical properties of ribosomes. New York: American Elsevier Publishing Co., Inc. 1964Google Scholar
  28. Rosenfeld, M. G., Abrass, I. B., Mendelsohn, J., Roos, B. A., Boone, R. F., Garren, L. D.: Control of transcription of RNA rich in polyadenylic acid in human lymphocytes. Proc. nat. Acad. Sci. (Wash.)69, 2306–2311 (1972)CrossRefGoogle Scholar
  29. 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.4, 1–10 (1956)PubMedGoogle Scholar
  30. Sheldon, R., Jurale, C., Kates, J.: Detection of polyadenylic acid sequences in viral and eukaryotic RNA. Proc. nat. Acad. Sci. (Wash.)69, 417–421 (1972)CrossRefGoogle Scholar
  31. 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-17β. Proc. nat. Acad. Sci. (Wash.)60, 1410–1417 (1968)CrossRefGoogle Scholar
  32. Thaler, M. M., Villee, C. A.: Template activities in normal, regenerating, and developing rat liver chromatin. Proc. nat. Acad. Sci. (Wash.)58, 2055–2062 (1967)CrossRefGoogle Scholar
  33. Weinberg, R. A.: Nuclear RNA Metabolism. Annu. Rev. Biochem.42, 329–354 (1973)PubMedCrossRefGoogle Scholar

Copyright information

© Springer-Verlag 1974

Authors and Affiliations

  • A. M. Novi
    • 1
    • 2
    • 3
  1. 1.Department of PathologyDüsseldorf UniversityDüsseldorgGermany
  2. 2.Department of Physiological Chemistry IDüsseldorf UniversityDüsseldorfGermany
  3. 3.Pathologisches Institut der UniversitätDüsseldorfFederal Republic of Germany

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