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PMID: 2657393 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

The phosphorylation state of eucaryotic initiation factor 2 alters translational efficiency of specific mRNAs.

Molecular and cellular biology ·Vol. 9 ·No. 3 ·1989-03-00 ·Pages 946-58

Kaufman RJ, Davies MV, Pathak VK, Hershey JW

Abstract

Phosphorylation of the alpha subunit of the eucaryotic translation initiation factor (eIF-2 alpha) by the double-stranded RNA-activated inhibitor (DAI) kinase correlates with inhibition of translation initiation. The importance of eIF-2 alpha phosphorylation in regulating translation was studied by expression of specific mutants of eIF-2 alpha in COS-1 cells. DNA transfection of certain plasmids could activate DAI kinase and result in poor translation of plasmid-derived mRNAs. In these cases, translation of the plasmid-derived mRNAs was improved by the presence of DAI kinase inhibitors or by the presence of a nonphosphorylatable mutant (serine to alanine) of eIF-2 alpha. The improved translation mediated by expression of the nonphosphorylatable eIF-2 alpha mutant was specific to plasmid-derived mRNA and did not affect global mRNA translation. Expression of a serine-to-aspartic acid mutant eIF-2 alpha, created to mimic the phosphorylated serine, inhibited translation of the mRNAs derived from the transfected plasmid. These results substantiate the hypothesis that DAI kinase activation reduces translation initiation through phosphorylation of eIF-2 alpha and reinforce the importance of phosphorylation of eIF-2 alpha as a way to control initiation of translation in intact cells.

MeSH Terms
Amino Acid Sequence Base Sequence DNA Probes Escherichia coli/genetics,metabolism Eukaryotic Initiation Factor-2 Humans Molecular Sequence Data Mutation Peptide Initiation Factors/genetics,metabolism Phosphorylation Plasmids Protein Biosynthesis Protein Kinases/metabolism Proteins/genetics,metabolism RNA, Messenger/genetics eIF-2 Kinase
Chemicals
DNA Probes Eukaryotic Initiation Factor-2 Peptide Initiation Factors Proteins RNA, Messenger Protein Kinases eIF-2 Kinase
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Kaufman R J
Genetics Institute, Cambridge, Massachusetts 02140.
Davies M V
Pathak V K
Hershey J W
References (50)
50 references, click to expand
  1. Selection and amplification of heterologous genes encoding adenosine deaminase in mammalian cells.
    Proc Natl Acad Sci U S A. 1986 May;83(10):3136-40 PMID: 3486414
  2. Purification of the eukaryotic initiation factor 2-eukaryotic initiation factor 2B complex and characterization of its guanine nucleotide exchange activity during protein synthesis initiation.
    J Biol Chem. 1983 Mar 10;258(5):3402-8 PMID: 6826566
  3. Initiation of protein synthesis in mammalian cells.
    Biochem J. 1986 May 1;235(3):625-37 PMID: 3530248
  4. Structure of pre-pro-von Willebrand factor and its expression in heterologous cells.
    Nature. 1986 Nov 20-26;324(6094):270-3 PMID: 3491324
  5. Regulation of polypeptide chain initiation in Chinese hamster ovary cells with a temperature-sensitive leucyl-tRNA synthetase. Changes in phosphorylation of initiation factor eIF-2 and in the activity of the guanine nucleotide exchange factor GEF.
    J Biol Chem. 1987 Jan 15;262(2):767-71 PMID: 3805006
  6. Cloning and sequencing of complementary DNAs encoding the alpha-subunit of translational initiation factor eIF-2. Characterization of the protein and its messenger RNA.
    J Biol Chem. 1987 Jan 25;262(3):1206-12 PMID: 2948954
  7. A mechanism by which adenovirus virus-associated RNAI controls translation in a transient expression assay.
    Mol Cell Biol. 1987 Jan;7(1):549-51 PMID: 2436038
  8. Initiation factor protein modifications and inhibition of protein synthesis.
    Mol Cell Biol. 1987 Mar;7(3):1293-5 PMID: 3561417
  9. Translational control mediated by eucaryotic initiation factor-2 is restricted to specific mRNAs in transfected cells.
    Mol Cell Biol. 1987 Apr;7(4):1568-71 PMID: 3600637
  10. Structure and regulation of eukaryotic initiation factor eIF-2. Sequence of the site in the alpha subunit phosphorylated by the haem-controlled repressor and by the double-stranded RNA-activated inhibitor.
    Eur J Biochem. 1987 Jul 15;166(2):357-63 PMID: 3609013
  11. Impact of virus infection on host cell protein synthesis.
    Annu Rev Biochem. 1987;56:317-32 PMID: 3113326
  12. Protein serine/threonine kinases.
    Annu Rev Biochem. 1987;56:567-613 PMID: 2956925
  13. Translational repression by chemical inducers of the stress response occurs by different pathways.
    Arch Biochem Biophys. 1987 Aug 1;256(2):651-61 PMID: 3619450
  14. Phosphorylation of eukaryotic initiation factor 2 during physiological stresses which affect protein synthesis.
    J Biol Chem. 1987 Oct 25;262(30):14538-43 PMID: 3667588
  15. Inhibition of rabbit reticulocyte lysate protein synthesis by heavy metal ions involves the phosphorylation of the alpha-subunit of the eukaryotic initiation factor 2.
    J Biol Chem. 1987 Nov 25;262(33):15939-45 PMID: 3680235
  16. Expression of a synthetic bovine rhodopsin gene in monkey kidney cells.
    Proc Natl Acad Sci U S A. 1987 Dec;84(24):8874-8 PMID: 2962193
  17. Three-dimensional structure of an oncogene protein: catalytic domain of human c-H-ras p21.
    Science. 1988 Feb 19;239(4842):888-93 PMID: 2448879
  18. Generation of a mutant form of protein synthesis initiation factor eIF-2 lacking the site of phosphorylation by eIF-2 kinases.
    Mol Cell Biol. 1988 Feb;8(2):993-5 PMID: 3352609
  19. The catalytic mechanism of guanine nucleotide exchange factor action and competitive inhibition by phosphorylated eukaryotic initiation factor 2.
    J Biol Chem. 1988 Apr 25;263(12):5526-33 PMID: 3356695
  20. Characterization of the double-stranded RNA implicated in the inhibition of protein synthesis in cells infected with a mutant adenovirus defective for VA RNA.
    Virology. 1988 May;164(1):106-13 PMID: 3363861
  21. Influenza virus regulates protein synthesis during infection by repressing autophosphorylation and activity of the cellular 68,000-Mr protein kinase.
    J Virol. 1988 Oct;62(10):3710-7 PMID: 3418783
  22. Cleavage of structural proteins during the assembly of the head of bacteriophage T4.
    Nature. 1970 Aug 15;227(5259):680-5 PMID: 5432063
  23. Synthesis of complementary RNA sequences during productive adenovirus infection.
    Proc Natl Acad Sci U S A. 1974 Dec;71(12):4887-91 PMID: 4612530
  24. DNA sequencing with chain-terminating inhibitors.
    Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463-7 PMID: 271968
  25. Quantitation of dihydrofolate reductase in individual parental and methotrexate-resistant murine cells. Use of a fluorescence activated cell sorter.
    J Biol Chem. 1978 Aug 25;253(16):5852-60 PMID: 670236
  26. Hybridization of denatured RNA and small DNA fragments transferred to nitrocellulose.
    Proc Natl Acad Sci U S A. 1980 Sep;77(9):5201-5 PMID: 6159641
  27. Transcriptional control in the production of liver-specific mRNAs.
    Cell. 1981 Mar;23(3):731-9 PMID: 7226226
  28. Amplification and expression of sequences cotransfected with a modular dihydrofolate reductase complementary dna gene.
    J Mol Biol. 1982 Aug 25;159(4):601-21 PMID: 6292436
  29. Adenovirus VAI RNA is required for efficient translation of viral mRNAs at late times after infection.
    Cell. 1982 Dec;31(3 Pt 2):543-51 PMID: 6297772
  30. Identification and quantitation of levels of protein synthesis initiation factors in crude HeLa cell lysates by two-dimensional polyacrylamide gel electrophoresis.
    J Biol Chem. 1983 Jun 10;258(11):7228-35 PMID: 6853516
  31. Regulation of protein synthesis initiation in eucaryotes.
    Arch Biochem Biophys. 1983 Jun;223(2):325-49 PMID: 6305278
  32. A GDP/GTP exchange factor essential for eukaryotic initiation factor 2 cycling in Ehrlich ascites tumor cells and its regulation by eukaryotic initiation factor 2 phosphorylation.
    J Biol Chem. 1983 Jul 10;258(13):7928-34 PMID: 6553052
  33. Mechanism of translational control by partial phosphorylation of the alpha subunit of eukaryotic initiation factor 2.
    Proc Natl Acad Sci U S A. 1984 Jan;81(2):352-6 PMID: 6320181
  34. Adenovirus VA RNAI: a positive regulator of mRNA translation.
    Mol Cell Biol. 1984 Apr;4(4):736-42 PMID: 6201722
  35. 2B or not 2B: regulation of the catalytic utilization of eIF-2.
    Cell. 1983 May;33(1):7-8 PMID: 6678613
  36. Inhibition of mRNA binding to ribosomes by localized activation of dsRNA-dependent protein kinase.
    Nature. 1984 Sep 6-11;311(5981):79-81 PMID: 6088993
  37. Heat shock-induced translational alterations in HeLa cells. Initiation factor modifications and the inhibition of translation.
    J Biol Chem. 1984 Oct 10;259(19):11882-9 PMID: 6384217
  38. Distribution of reversing factor in reticulocyte lysates during active protein synthesis and on inhibition by heme deprivation or double-stranded RNA.
    Proc Natl Acad Sci U S A. 1984 Nov;81(22):6998-7002 PMID: 6594676
  39. Regulation of a protein synthesis initiation factor by adenovirus virus-associated RNA.
    Nature. 1985 Jan 17-23;313(5999):196-200 PMID: 2578613
  40. Identification of the components necessary for adenovirus translational control and their utilization in cDNA expression vectors.
    Proc Natl Acad Sci U S A. 1985 Feb;82(3):689-93 PMID: 2983309
  41. 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):1959-63 PMID: 3856874
  42. Regulation of initiation factors during translational repression caused by serum depletion. Covalent modification.
    J Biol Chem. 1985 May 10;260(9):5493-7 PMID: 3886657
  43. Double-stranded RNA-dependent protein kinase and 2-5A system are both activated in interferon-treated, encephalomyocarditis virus-infected HeLa cells.
    J Virol. 1985 Jun;54(3):894-8 PMID: 2582150
  44. Human GM-CSF: molecular cloning of the complementary DNA and purification of the natural and recombinant proteins.
    Science. 1985 May 17;228(4701):810-5 PMID: 3923623
  45. 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):4321-5 PMID: 2989814
  46. Adenovirus VA RNAI mediates a translational stimulation which is not restricted to the viral mRNAs.
    EMBO J. 1985 Apr;4(4):957-64 PMID: 2410262
  47. Activation of hemin-regulated initiation factor-2 kinase in heat-shocked HeLa cells.
    J Biol Chem. 1986 Jan 5;261(1):338-42 PMID: 3941080
  48. A mechanism for the control of protein synthesis by adenovirus VA RNAI.
    Cell. 1986 Feb 14;44(3):391-400 PMID: 3943131
  49. Adenovirus VAI RNA antagonizes the antiviral action of interferon by preventing activation of the interferon-induced eIF-2 alpha kinase.
    Cell. 1986 Apr 25;45(2):195-200 PMID: 3698097
  50. The NH2-terminal sequence of the alpha and gamma subunits of eukaryotic initiation factor 2 and the phosphorylation site for the heme-regulated eIF-2 alpha kinase.
    J Biol Chem. 1986 Sep 25;261(27):12444-7 PMID: 3745199
Article Info
Journal
Molecular and cellular biology
Abbr.
Mol Cell Biol
ISSN
0270-7306
DOI
10.1128/mcb.9.3.946-958.1989
Published
1989-03-00
Pages
946-58
Language
English
Region
United States
NLM ID
8109087
PMCID
PMC362683
Subset
IM
Grants
Wellcome Trust · 089701 · United Kingdom
NIGMS NIH HHS · GM22135 · United States
External Links
PubMed source DOI: 10.1128/mcb.9.3.946-958.1989 PMC Full Text View journal details
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