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. 1986 Dec;60(3):1012–1017. doi: 10.1128/jvi.60.3.1012-1017.1986

A kinase able to phosphorylate exogenous protein synthesis initiation factor eIF-2 alpha is present in lysates of mengovirus-infected L cells.

A Pani, M Julian, J Lucas-Lenard
PMCID: PMC253340  PMID: 3023653

Abstract

Infection of mouse L929 cells by mengovirus resulted in the expression of a kinase activity that selectively phosphorylated the small, 38,000-molecular-weight subunit of eucaryotic initiation factor 2 and histone H2. This kinase activity was independent of host cell RNA synthesis and was located in the postribosomal supernatant (S-100 fraction) early after infection (up to 3 h). At later times after infection (5 h), kinase activity was also associated with the polysome fraction. The kinase present in the S-100 fraction bound strongly to DEAE-cellulose, its peak activity eluting at 0.5 M KCl. Kinase activity was independent of the presence of exogenous double-stranded RNA, and KCl at concentrations greater than 0.1 M inhibited eucaryotic initiation factor 2 phosphorylation. The 67,000-molecular-weight phosphoprotein activated in interferon-treated cells by double-stranded RNA was not detected by standard phosphorylation assays in lysates from mengovirus-infected cells. Labeling of this protein in vivo during 5 h of infection was also not detected. The DEAE-cellulose-purified mengovirus kinase inhibited protein synthesis in reticulocyte lysates, and the inhibition was not reversible by high concentrations of poly(I).poly(C).

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Selected References

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

  1. Abreu S. L., Lucas-Lenard J. Cellular protein synthesis shutoff by mengovirus: translation of nonviral and viral mRNA's in extracts from uninfected and infected Ehrlich ascites tumor cells. J Virol. 1976 Apr;18(1):182–194. doi: 10.1128/jvi.18.1.182-194.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Berry M. J., Knutson G. S., Lasky S. R., Munemitsu S. M., Samuel C. E. Mechanism of interferon action. Purification and substrate specificities of the double-stranded RNA-dependent protein kinase from untreated and interferon-treated mouse fibroblasts. J Biol Chem. 1985 Sep 15;260(20):11240–11247. [PubMed] [Google Scholar]
  3. Brown G. D., Peluso R. W., Moyer S. A., Moyer R. W. A simple method for the preparation of extracts from animal cells which catalyze efficient in vitro protein synthesis. J Biol Chem. 1983 Dec 10;258(23):14309–14314. [PubMed] [Google Scholar]
  4. Das H. K., Das A., Ghosh-Dastidar P., Ralston R. O., Yaghmai B., Roy R., Gupta N. K. Protein synthesis in rabbit reticulocytes. Purification and characterization of a double-stranded RNA-dependent protein synthesis inhibitor from reticulocyte lysates. J Biol Chem. 1981 Jun 25;256(12):6491–6495. [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. Specific protein phosphorylation in interferon-treated uninfected and virus-infected mouse L929 cells: enhancement by double-stranded RNA. J Virol. 1979 Jan;29(1):301–311. doi: 10.1128/jvi.29.1.301-311.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Hunter T., Hunt T., Jackson R. J., Robertson H. D. The characteristics of inhibition of protein synthesis by double-stranded ribonucleic acid in reticulocyte lysates. J Biol Chem. 1975 Jan 25;250(2):409–417. [PubMed] [Google Scholar]
  8. Jaye M. C., Godchaux W., 3rd, Lucas-Lenard J. Further studies on the inhibition of cellular protein synthesis by vesicular stomatitis virus. Virology. 1982 Jan 15;116(1):148–162. doi: 10.1016/0042-6822(82)90410-x. [DOI] [PubMed] [Google Scholar]
  9. Kimchi A., Zilberstein A., Schmidt A., Shulman L., Revel M. The interferon-induced protein kinase PK-i from mouse L cells. J Biol Chem. 1979 Oct 10;254(19):9846–9853. [PubMed] [Google Scholar]
  10. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  11. Levin D. H., Petryshyn R., London I. M. Characterization of double-stranded-RNA-activated kinase that phosphorylates alpha subunit of eukaryotic initiation factor 2 (eIF-2 alpha) in reticulocyte lysates. Proc Natl Acad Sci U S A. 1980 Feb;77(2):832–836. doi: 10.1073/pnas.77.2.832. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Levin D. H., Petryshyn R., London I. M. Characterization of purified double-stranded RNA-activated eIF-2 alpha kinase from rabbit reticulocytes. J Biol Chem. 1981 Jul 25;256(14):7638–7641. [PubMed] [Google Scholar]
  13. Marcus P. I., Guidon P. T., Jr, Sekellick M. J. Interferon induction by viruses. VII. Mengovirus: "interferon-sensitive" mutant phenotype attributed to interferon-inducing particle activity. J Interferon Res. 1981;1(4):601–611. doi: 10.1089/jir.1981.1.601. [DOI] [PubMed] [Google Scholar]
  14. Ochoa S. Regulation of protein synthesis initiation in eucaryotes. Arch Biochem Biophys. 1983 Jun;223(2):325–349. doi: 10.1016/0003-9861(83)90598-2. [DOI] [PubMed] [Google Scholar]
  15. Paez E., Esteban M. Resistance of vaccinia virus to interferon is related to an interference phenomenon between the virus and the interferon system. Virology. 1984 Apr 15;134(1):12–28. doi: 10.1016/0042-6822(84)90268-x. [DOI] [PubMed] [Google Scholar]
  16. Pensiero M. N., Lucas-Lenard J. M. Evidence for the presence of an inhibitor on ribosomes in mouse L cells infected with mengovirus. J Virol. 1985 Oct;56(1):161–171. doi: 10.1128/jvi.56.1.161-171.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Rice A. P., Kerr I. M. Interferon-mediated, double-stranded RNA-dependent protein kinase is inhibited in extracts from vaccinia virus-infected cells. J Virol. 1984 Apr;50(1):229–236. doi: 10.1128/jvi.50.1.229-236.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Rosen H., Knoller S., Kaempfer R. Messenger ribonucleic acid specificity in the inhibition of eukaryotic translation by double-stranded ribonucleic acid. Biochemistry. 1981 May 26;20(11):3011–3020. doi: 10.1021/bi00514a004. [DOI] [PubMed] [Google Scholar]
  19. Rosnitschek I., Traub U., Traub P. Enhanced phosphorylation of ribosomal protein s6 and other cytoplasmic proteins after mengovirus infection of Ehrlich ascites tumor cells. Hoppe Seylers Z Physiol Chem. 1978 May;359(5):593–600. doi: 10.1515/bchm.1978.359.1.593. [DOI] [PubMed] [Google Scholar]
  20. Samuel C. E., Duncan R., Knutson G. S., Hershey J. W. Mechanism of interferon action. Increased phosphorylation of protein synthesis initiation factor eIF-2 alpha in interferon-treated, reovirus-infected mouse L929 fibroblasts in vitro and in vivo. J Biol Chem. 1984 Nov 10;259(21):13451–13457. [PubMed] [Google Scholar]
  21. Schneider R. J., Safer B., Munemitsu S. M., Samuel C. E., Shenk T. Adenovirus VAI RNA prevents phosphorylation of the eukaryotic initiation factor 2 alpha subunit subsequent to infection. Proc Natl Acad Sci U S A. 1985 Jul;82(13):4321–4325. doi: 10.1073/pnas.82.13.4321. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Sen G. C., Taira H., Lengyel P. Interferon, double-stranded RNA, and protein phosphorylation. Characteristics of a double-stranded RNA-activated protein kinase system partially purified from interferon treated Ehrlich ascites tumor cells. J Biol Chem. 1978 Sep 10;253(17):5915–5921. [PubMed] [Google Scholar]
  23. Siekierka J., Mariano T. M., Reichel P. A., Mathews M. B. Translational control by adenovirus: lack of virus-associated RNAI during adenovirus infection results in phosphorylation of initiation factor eIF-2 and inhibition of protein synthesis. Proc Natl Acad Sci U S A. 1985 Apr;82(7):1959–1963. doi: 10.1073/pnas.82.7.1959. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Simon E. H., Kung S., Koh T. T., Brandman P. Interferon-sensitive mutants of mengovirus. I. Isolation and biological characterization. Virology. 1976 Feb;69(2):727–736. doi: 10.1016/0042-6822(76)90501-8. [DOI] [PubMed] [Google Scholar]
  25. Tahara S. M., Traugh J. A., Sharp S. B., Lundak T. S., Safer B., Merrick W. C. Effect of hemin on site-specific phosphorylation of eukaryotic initiation factor 2. Proc Natl Acad Sci U S A. 1978 Feb;75(2):789–793. doi: 10.1073/pnas.75.2.789. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Tuazon P. T., Merrick W. C., Traugh J. A. Site-specific phosphorylation if initiation factor 2 by three cyclic nucleotide-independent protein kinases. J Biol Chem. 1980 Nov 25;255(22):10954–10958. [PubMed] [Google Scholar]
  27. Whitaker-Dowling P., Youngner J. S. Characterization of a specific kinase inhibitory factor produced by vaccinia virus which inhibits the interferon-induced protein kinase. Virology. 1984 Aug;137(1):171–181. doi: 10.1016/0042-6822(84)90020-5. [DOI] [PubMed] [Google Scholar]

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