Skip to main content
PMC home page
Molecular and Cellular Biology logoLink to Molecular and Cellular Biology
. 1981 Oct;1(10):932–938. doi: 10.1128/mcb.1.10.932

Elevated levels of (2'-5')oligoadenylic acid polymerase activity in growth-arrested human lymphoblastoid Namalva cells.

I Krishnan, C Baglioni
PMCID: PMC369381  PMID: 6180294

Abstract

(2'-5')Oligoadenylic acid [(2'-5')An] polymerase activity was measured in extracts of human lymphoblastoid cells of the Namalva line cultured under different conditions. Exponentially growing cells had a relatively low polymerase activity level, whereas cells grown to limit density showed elevated levels. When fresh medium was added to growth-arrested cells, (2'-5')An polymerase activity decreased concomitantly with the initiation of active deoxyribonucleic acid synthesis. An increase in polymerase activity level was also observed after exponentially growing cells were transferred from medium containing 20% serum to fresh medium containing 0.2% serum. These cells diminished deoxyribonucleic acid synthesis and remained quiescent until 20% serum was again added. Polymerase activity level decreased as the cells entered into S phase. The addition of the inhibitor of deoxyribonucleic acid synthesis, hydroxyurea, to exponentially growing cells did not increase polymerase level, indicating that cells blocked in S phase and at the G1-S boundary maintained the basal level of this enzyme. Degradation of labeled (2'-5')An was measured in extracts of Namalva cells cultured under different conditions, but no significant differences among degradative activities were observed. Since (2'-5')An polymerase activity is one of the enzymatic activities induced by interferon, we measured interferon titers in Namalva cell medium. Less than 1 reference unit per ml was detected in cells grown under different conditions. Moreover, the increase in (2'-5')An polymerase activity level in cells transferred from 20 to 0.2% serum was not prevented by including anti-lymphoblastoid interferon antibody in the medium. These results suggest that the activity level of (2'-5')An polymerase is regulated in Namalva cells on the basis of the growth status of the cells and that this regulatory mechanism is apparently not activated by interferon.

Full text

PDF
932

Selected References

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

  1. Baglioni C. Interferon-induced enzymatic activities and their role in the antriviral state. Cell. 1979 Jun;17(2):255–264. doi: 10.1016/0092-8674(79)90151-x. [DOI] [PubMed] [Google Scholar]
  2. Baglioni C., Minks M. A., Maroney P. A. Interferon action may be mediated by activation of a nuclease by pppA2'p5'A2'p5'A. Nature. 1978 Jun 22;273(5664):684–687. doi: 10.1038/273684a0. [DOI] [PubMed] [Google Scholar]
  3. Borden E. C., Leonhardt P. H. Aquanititave semimicro, semiautomated colorimetric assay for interferon. J Lab Clin Med. 1977 May;89(5):1036–1042. [PubMed] [Google Scholar]
  4. Clemens M. J., Williams B. R. Inhibition of cell-free protein synthesis by pppA2'p5'A2'p5'A: a novel oligonucleotide synthesized by interferon-treated L cell extracts. Cell. 1978 Mar;13(3):565–572. doi: 10.1016/0092-8674(78)90329-x. [DOI] [PubMed] [Google Scholar]
  5. Farrell P. J., Balkow K., Hunt T., Jackson R. J., Trachsel H. Phosphorylation of initiation factor elF-2 and the control of reticulocyte protein synthesis. Cell. 1977 May;11(1):187–200. doi: 10.1016/0092-8674(77)90330-0. [DOI] [PubMed] [Google Scholar]
  6. Gupta S. L., Rubin B. Y., Holmes S. L. Interferon action: induction of specific proteins in mouse and human cells by homologous interferons. Proc Natl Acad Sci U S A. 1979 Oct;76(10):4817–4821. doi: 10.1073/pnas.76.10.4817. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Hovanessian A. G., Kerr I. M. Synthesis of an oligonucleotide inhibitor of protein synthesis in rabbit reticulocyte lysates analogous to that formed in extracts from interferon-treated cells. Eur J Biochem. 1978 Mar;84(1):149–159. doi: 10.1111/j.1432-1033.1978.tb12151.x. [DOI] [PubMed] [Google Scholar]
  8. Hovanessian A. G., Wood J., Meurs E., Montagnier L. Increased nuclease activity in cells treated with pppA2'p5'A2'p5' A. Proc Natl Acad Sci U S A. 1979 Jul;76(7):3261–3265. doi: 10.1073/pnas.76.7.3261. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Kerr I. M., Brown R. E. pppA2'p5'A2'p5'A: an inhibitor of protein synthesis synthesized with an enzyme fraction from interferon-treated cells. Proc Natl Acad Sci U S A. 1978 Jan;75(1):256–260. doi: 10.1073/pnas.75.1.256. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Kimchi A., Shure H., Revel M. Regulation of lymphocyte mitogenesis by (2'--5') oligo-isoadenylate. Nature. 1979 Dec 20;282(5741):849–851. doi: 10.1038/282849a0. [DOI] [PubMed] [Google Scholar]
  11. Knight E., Jr, Korant B. D. Fibroblast interferon induces synthesis of four proteins in human fibroblast cells. Proc Natl Acad Sci U S A. 1979 Apr;76(4):1824–1827. doi: 10.1073/pnas.76.4.1824. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Knight M., Cayley P. J., Silverman R. H., Wreschner D. H., Gilbert C. S., Brown R. E., Kerr I. M. Radioimmune, radiobinding and HPLC analysis of 2-5A and related oligonucleotides from intact cells. Nature. 1980 Nov 13;288(5787):189–192. doi: 10.1038/288189a0. [DOI] [PubMed] [Google Scholar]
  13. Krishnan I., Baglioni C. Increased levels of (2'-5')oligo(A) polymerase activity in human lymphoblastoid cells treated with glucocorticoids. Proc Natl Acad Sci U S A. 1980 Nov;77(11):6506–6510. doi: 10.1073/pnas.77.11.6506. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Krishnan I., Baglioni C. Regulation of 2'5'-oligo(A) polymerase activity in quiescent human fibroblasts treated with interferon. Virology. 1981 Jun;111(2):666–670. doi: 10.1016/0042-6822(81)90368-8. [DOI] [PubMed] [Google Scholar]
  15. Minks M. A., Benvin S., Maroney P. A., Baglioni C. Metabolic stability of 2' 5'oligo (A) and activity of 2' 5'oligo (A)-dependent endonuclease in extracts of interferon-treated and control HeLa cells. Nucleic Acids Res. 1979 Feb;6(2):767–780. doi: 10.1093/nar/6.2.767. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Minks M. A., Benvin S., Maroney P. A., Baglioni C. Synthesis of 2'5'-oligo(A) in extracts of interferon-treated HeLa cells. J Biol Chem. 1979 Jun 25;254(12):5058–5064. [PubMed] [Google Scholar]
  17. Pickering L. A., Kronenberg L. H., Stewart W. E., 2nd Spontaneous production of human interferon. Proc Natl Acad Sci U S A. 1980 Oct;77(10):5938–5942. doi: 10.1073/pnas.77.10.5938. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Ratner L., Wiegand R. C., Farrell P. J., Sen G. C., Cabrer B., Lengyel P. Interferon, double-stranded RNA and RNA degradation. Fractionation of the endonucleaseINT system into two macromolecular components; role of a small molecule in nuclease activation. Biochem Biophys Res Commun. 1978 Apr 14;81(3):947–954. doi: 10.1016/0006-291x(78)91443-2. [DOI] [PubMed] [Google Scholar]
  19. Schmidt A., Chernajovsky Y., Shulman L., Federman P., Berissi H., Revel M. An interferon-induced phosphodiesterase degrading (2'-5') oligoisoadenylate and the C-C-A terminus of tRNA. Proc Natl Acad Sci U S A. 1979 Oct;76(10):4788–4792. doi: 10.1073/pnas.76.10.4788. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Shimizu N., Sokawa Y. 2',5'-Oligoadenylate synthetase activity in lymphocytes from normal mouse. J Biol Chem. 1979 Dec 10;254(23):12034–12037. [PubMed] [Google Scholar]
  21. Shulman L., Revel M. Interferon-dependent induction of mRNA activity for (2'-5')oligo-isoadenylate synthetase. Nature. 1980 Nov 6;288(5786):98–100. doi: 10.1038/288098a0. [DOI] [PubMed] [Google Scholar]
  22. Walters R. A., Tobey R. A., Hildebrand C. E. Hydroxyurea does not prevent synchronized G1 chinese hamster cells from entering the DNA synthetic period. Biochem Biophys Res Commun. 1976 Mar 8;69(1):212–217. doi: 10.1016/s0006-291x(76)80294-x. [DOI] [PubMed] [Google Scholar]
  23. Zajac B. A., Henle W., Henle G. Autogenous and virus-induced interferons from lines of lymphoblastoid cells. Cancer Res. 1969 Aug;29(8):1467–1475. [PubMed] [Google Scholar]
  24. Zoon K. C., Smith M. E., Bridgen P. J., zur Nedden D., Anfinsen C. B. Purification and partial characterization of human lymphoblast interferon. Proc Natl Acad Sci U S A. 1979 Nov;76(11):5601–5605. doi: 10.1073/pnas.76.11.5601. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Molecular and Cellular Biology are provided here courtesy of Taylor & Francis

RESOURCES