Abstract
We recently reported that interferon induces the synthesis of ppp(A2'p)nA(n = 2 to greater than or equal to 4) (2-5A)-dependent RNase in the murine cell line JLS-V9R. These cells normally contain very low levels of the nuclease; after interferon treatment, however, they develop levels approaching those found in murine L or Ehrlich ascites tumor cells. Here, we report a similar increase in the nuclease levels in JLS-V9R cells during the transition from the subconfluent actively growing state to the confluent stationary phase. Levels of 2-5A synthetase increased in parallel with the nuclease. The induced levels of both the nuclease and synthetase returned to low basal amounts after trypsinization, dilution, and culturing of the cells at subconfluent densities. The addition of anti-murine interferon (alpha + beta) antibodies to the medium did not affect the induction of the nuclease nor could any interferon be detected in the culture supernatants as determined by the lack of antiviral activity. The increase in the enzymes was not, therefore, due to the spontaneous production of interferon. The induction of the nuclease during confluency preceded an inhibition of [3H]-thymidine incorporation by the cells into DNA. The regulation of the 2-5A-dependent RNase in JLS-V9R cells may, therefore, be related to the control of cell growth.
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- Adams A., Lidin B., Strander H., Cantell K. Spontaneous interferon production and Epstein-Barr virus antigen expression in human lymphoid cell lines. J Gen Virol. 1975 Aug;28(2):219–223. doi: 10.1099/0022-1317-28-2-219. [DOI] [PubMed] [Google Scholar]
- Allen P. T., Schellekens H., Van Griensven L. J., Billiau A. Differential sensitivity of Rauscher murine leukaemia virus (MuLV-R) to interferons in two interferon-responsive cell lines. J Gen Virol. 1976 Jun;31(3):429–435. doi: 10.1099/0022-1317-31-3-429. [DOI] [PubMed] [Google Scholar]
- 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]
- Ball L. A. Induction of 2'5'-oligoadenylate synthetase activity and a new protein by chick interferon. Virology. 1979 Apr 30;94(2):282–296. doi: 10.1016/0042-6822(79)90462-8. [DOI] [PubMed] [Google Scholar]
- Bradford M. M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976 May 7;72:248–254. doi: 10.1006/abio.1976.9999. [DOI] [PubMed] [Google Scholar]
- Cayley P. J., Knight M., Kerr I. M. Virus-mediated inhibition of the ppp(A2'p)nA system and its prevention by interferon. Biochem Biophys Res Commun. 1982 Jan 29;104(2):376–382. doi: 10.1016/0006-291x(82)90647-7. [DOI] [PubMed] [Google Scholar]
- 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]
- Floyd-Smith G., Yoshie O., Lengyel P. Interferon action. Covalent linkage of (2'-5')pppApApA(32P)pCp to (2'-5')(A)n-dependent ribonucleases in cell extracts by ultraviolet irradiation. J Biol Chem. 1982 Aug 10;257(15):8584–8587. [PubMed] [Google Scholar]
- Hovanessian A. G., Wood J. N. Anticellular and antiviral effects of pppA(2'p5'A)n. Virology. 1980 Feb;101(1):81–90. doi: 10.1016/0042-6822(80)90485-7. [DOI] [PubMed] [Google Scholar]
- 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]
- Imai J., Johnston M. I., Torrence P. F. Chemical modification potentiates the biological activities of 2-5A and its congeners. J Biol Chem. 1982 Nov 10;257(21):12739–12745. [PubMed] [Google Scholar]
- Jacobsen H., Czarniecki C. W., Krause D., Friedman R. M., Silverman R. H. Interferon-induced synthesis of 2-5A-dependent RNase in mouse JLS-V9R cells. Virology. 1983 Mar;125(2):496–501. doi: 10.1016/0042-6822(83)90222-2. [DOI] [PubMed] [Google Scholar]
- Kerr I. M., Brown R. E., Hovanessian A. G. Nature of inhibitor of cell-free protein synthesis formed in response to interferon and double-stranded RNA. Nature. 1977 Aug 11;268(5620):540–542. doi: 10.1038/268540a0. [DOI] [PubMed] [Google Scholar]
- 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]
- Kimchi A., Shulman L., Schmidt A., Chernajovsky Y., Fradin A., Revel M. Kinetics of the induction of three translation-regulatory enzymes by interferon. Proc Natl Acad Sci U S A. 1979 Jul;76(7):3208–3212. doi: 10.1073/pnas.76.7.3208. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 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]
- Lebleu B., Sen G. C., Shaila S., Cabrer B., Lengyel P. Interferon, double-stranded RNA, and protein phosphorylation. Proc Natl Acad Sci U S A. 1976 Sep;73(9):3107–3111. doi: 10.1073/pnas.73.9.3107. [DOI] [PMC free article] [PubMed] [Google Scholar]
- McMaster G. K., Carmichael G. G. Analysis of single- and double-stranded nucleic acids on polyacrylamide and agarose gels by using glyoxal and acridine orange. Proc Natl Acad Sci U S A. 1977 Nov;74(11):4835–4838. doi: 10.1073/pnas.74.11.4835. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 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]
- Nilsen T. W., McCandless S., Baglioni C. 2', 5'-oligo(A)-activated endonuclease in NIH 3T3 mouse cells chronically infected with Moloney murine leukemia virus. Virology. 1982 Oct 30;122(2):498–502. doi: 10.1016/0042-6822(82)90252-5. [DOI] [PubMed] [Google Scholar]
- Nilsen T. W., Wood D. L., Baglioni C. 2',5'-Oligo(A)-activated endoribonuclease. Tissue distribution and characterization with a binding assay. J Biol Chem. 1981 Nov 10;256(21):10751–10754. [PubMed] [Google Scholar]
- 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]
- Roberts W. K., Hovanessian A., Brown R. E., Clemens M. J., Kerr I. M. Interferon-mediated protein kinase and low-molecular-weight inhibitor of protein synthesis. Nature. 1976 Dec 2;264(5585):477–480. doi: 10.1038/264477a0. [DOI] [PubMed] [Google Scholar]
- 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]
- Silverman R. H., Cayley P. J., Knight M., Gilbert C. S., Kerr I. M. Control of the ppp(a2'p)nA system in HeLa cells. Effects of interferon and virus infection. Eur J Biochem. 1982 May;124(1):131–138. doi: 10.1111/j.1432-1033.1982.tb05915.x. [DOI] [PubMed] [Google Scholar]
- Silverman R. H., Watling D., Balkwill F. R., Trowsdale J., Kerr I. M. The ppp(A2'p)nA and protein kinase systems in wild-type and interferon-resistant Daudi cells. Eur J Biochem. 1982 Aug;126(2):333–341. doi: 10.1111/j.1432-1033.1982.tb06783.x. [DOI] [PubMed] [Google Scholar]
- Silverman R. H., Wreschner D. H., Gilbert C. S., Kerr I. M. Synthesis, characterization and properties of ppp(A2'p)nApCp and related high-specific-activity 32P-labelled derivatives of ppp(A2'p)nA. Eur J Biochem. 1981 Mar 16;115(1):79–85. doi: 10.1111/j.1432-1033.1981.tb06200.x. [DOI] [PubMed] [Google Scholar]
- Stark G. R., Brown R. E., Kerr I. M. Assay of (2'-5')-oligoadenylic acid synthetase levels in cells and tissues: a convenient poly(I) . poly(C) paper-bound enzyme assay. Methods Enzymol. 1981;79(Pt B):194–199. doi: 10.1016/s0076-6879(81)79029-3. [DOI] [PubMed] [Google Scholar]
- Stark G. R., Dower W. J., Schimke R. T., Brown R. E., Kerr I. M. 2-5A synthetase: assay, distribution and variation with growth or hormone status. Nature. 1979 Mar 29;278(5703):471–473. doi: 10.1038/278471a0. [DOI] [PubMed] [Google Scholar]
- Verhaegen-Lewalle M., Content J. 2'-Phosphodiesterase activity in human cell lines treated or untreated with human interferon. Eur J Biochem. 1982 Sep 1;126(3):639–643. doi: 10.1111/j.1432-1033.1982.tb06828.x. [DOI] [PubMed] [Google Scholar]
- Williams B. R., Kerr I. M., Gilbert C. S., White C. N., Ball L. A. Synthesis and breakdown of pppA2'p5'A2'p5'A and transient inhibiton of protein synthesis in extracts from interferon-treated and control cells. Eur J Biochem. 1978 Dec;92(2):455–462. doi: 10.1111/j.1432-1033.1978.tb12767.x. [DOI] [PubMed] [Google Scholar]
- Williams B. R., Kerr I. M. Inhibition of protein synthesis by 2'-5' linked adenine oligonucleotides in intact cells. Nature. 1978 Nov 2;276(5683):88–90. doi: 10.1038/276088a0. [DOI] [PubMed] [Google Scholar]
- Wreschner D. H., James T. C., Silverman R. H., Kerr I. M. Ribosomal RNA cleavage, nuclease activation and 2-5A(ppp(A2'p)nA) in interferon-treated cells. Nucleic Acids Res. 1981 Apr 10;9(7):1571–1581. doi: 10.1093/nar/9.7.1571. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Wreschner D. H., Silverman R. H., James T. C., Gilbert C. S., Kerr I. M. Affinity labelling and characterization of the ppp(A2'p)nA-dependent endoribonuclease from different mammalian sources. Eur J Biochem. 1982 May 17;124(2):261–268. doi: 10.1111/j.1432-1033.1982.tb06586.x. [DOI] [PubMed] [Google Scholar]
- Zilberstein A., Federman P., Shulman L., Revel M. Specific phosphorylation in vitro of a protein associated with ribosomes of interferon-treated mouse L cells. FEBS Lett. 1976 Sep 15;68(1):119–124. doi: 10.1016/0014-5793(76)80418-8. [DOI] [PubMed] [Google Scholar]