Purification and Properties of the Double-stranded RNA-activated Eukaryotic Initiation Factor 3 Kinase from Rabbit Reticulocytes

Overview

Journal Proc Natl Acad Sci U S A

Specialty Science

Date 1980 Nov 1

PMID 6935666

Citations 7

Authors

H Grosfeld

S Ochoa

Affiliations

Soon will be listed here.

Abstract

The double-stranded RNA (dsRNA)-activated protein kinase (DAI) that phosphorylates the alpha subunit of the eukaryotic initiation factor eIF-2 and inhibits chain initiation has been isolated from rabbit reticulocyte lysates. The nonactivated enzyme or the enzyme partially activated by incubation with low levels of dsRNA (pro-DAI) could be purified only to a slight extent. However, the enzyme that was fully activated by incubation with both dsRNA and ATP was purified to near homogeneity. Active DAI is a phosphoprotein with an apparent subunit mass of 68,000 daltons. It can phosphorylate histone as well as the alpha subunit of eIF-2. Our results suggest that, after interaction with dsRNA, the enzyme phosphorylates itself and is thereby activated to phosphorylate alpha eIF-2 and histone.

Citing Articles

Protein synthesis in rabbit reticulocytes: characteristics of the protein factor RF that reverses inhibition of protein synthesis in heme-deficient reticulocyte lysates.

Grace M, Ralston R, Banerjee A, Gupta N Proc Natl Acad Sci U S A. 1982; 79(21):6517-21.

PMID: 6959132 PMC: 347158. DOI: 10.1073/pnas.79.21.6517.

Mode of action of the heme-controlled translational inhibitor: relationship of eukaryotic initiation factor 2-stimulating protein to translation restoring factor.

Siekierka J, Mitsui K, Ochoa S Proc Natl Acad Sci U S A. 1981; 78(1):220-3.

PMID: 6941245 PMC: 319023. DOI: 10.1073/pnas.78.1.220.

Regulation of double-stranded RNA-activated eukaryotic initiation factor 2 alpha kinase by type 2 protein phosphatase in reticulocyte lysates.

Petryshyn R, LEVIN D, LONDON I Proc Natl Acad Sci U S A. 1982; 79(21):6512-6.

PMID: 6292906 PMC: 347157. DOI: 10.1073/pnas.79.21.6512.

Regulation of protein synthesis in vesicular stomatitis virus-infected mouse L-929 cells by decreased protein synthesis initiation factor 2 activity.

Centrella M J Virol. 1982; 41(3):781-91.

PMID: 6284970 PMC: 256815. DOI: 10.1128/JVI.41.3.781-791.1982.

Double-stranded RNA-dependent phosphorylation of protein P1 and eukaryotic initiation factor 2 alpha does not correlate with protein synthesis inhibition in a cell-free system from interferon-treated mouse L cells.

Jacobsen H, Epstein D, Friedmann R, Safer B, Torrence P Proc Natl Acad Sci U S A. 1983; 80(1):41-5.

PMID: 6185959 PMC: 393305. DOI: 10.1073/pnas.80.1.41.

References

Laemmli U . Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970; 227(5259):680-5. DOI: 10.1038/227680a0. View

Gross M, Rabinovitz M . Control of globin synthesis by hemin: factors influencing formation of an inhibitor of globin chain initiation in reticulocyte lysates. Biochim Biophys Acta. 1972; 287(2):340-52. DOI: 10.1016/0005-2787(72)90383-8. View

Bradford M . A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976; 72:248-54. DOI: 10.1016/0003-2697(76)90527-3. View

Kramer G, Cimadevilla J, Hardesty B . Specificity of the protein kinase activity associated with the hemin-controlled repressor of rabbit reticulocyte. Proc Natl Acad Sci U S A. 1976; 73(9):3078-82. PMC: 430935. DOI: 10.1073/pnas.73.9.3078. View

Levin D, Ranu R, Ernst V, LONDON I . Regulation of protein synthesis in reticulocyte lysates: phosphorylation of methionyl-tRNAf binding factor by protein kinase activity of translational inhibitor isolated from hemedeficient lysates. Proc Natl Acad Sci U S A. 1976; 73(9):3112-6. PMC: 430947. DOI: 10.1073/pnas.73.9.3112. View

Gross M, Mendelewski J . Additional evidence that the hemin-controlled translational repressor from rabbit reticulocytes is a protein kinase. Biochem Biophys Res Commun. 1977; 74(2):559-69. DOI: 10.1016/0006-291x(77)90340-0. View

Farrell P, Balkow K, Hunt T, Jackson R, Trachsel H . Phosphorylation of initiation factor elF-2 and the control of reticulocyte protein synthesis. Cell. 1977; 11(1):187-200. DOI: 10.1016/0092-8674(77)90330-0. View

Benne R, Edman J, Traut R, Hershey J . Phosphorylation of eukaryotic protein synthesis initiation factors. Proc Natl Acad Sci U S A. 1978; 75(1):108-12. PMC: 411193. DOI: 10.1073/pnas.75.1.108. View

Kerr I, Brown R . 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; 75(1):256-60. PMC: 411225. DOI: 10.1073/pnas.75.1.256. View

10.

Tahara S, Traugh J, Sharp S, Lundak T, Safer B, Merrick W . Effect of hemin on site-specific phosphorylation of eukaryotic initiation factor 2. Proc Natl Acad Sci U S A. 1978; 75(2):789-93. PMC: 411342. DOI: 10.1073/pnas.75.2.789. View

11.

Levin D, LONDON I . Regulation of protein synthesis: activation by double-stranded RNA of a protein kinase that phosphorylates eukaryotic initiation factor 2. Proc Natl Acad Sci U S A. 1978; 75(3):1121-5. PMC: 411420. DOI: 10.1073/pnas.75.3.1121. View

12.

Lenz J, Baglioni C . Inhibition of protein synthesis by double-stranded RNA and phosphorylation of initiation factor, eIF-2. J Biol Chem. 1978; 253(12):4219-23. View

13.

de Haro C, Ochoa S . Mode of action of the hemin-controlled inhibitor of protein synthesis: studies with factors from rabbit reticulocytes. Proc Natl Acad Sci U S A. 1978; 75(6):2713-6. PMC: 392633. DOI: 10.1073/pnas.75.6.2713. View

14.

Sen G, 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; 253(17):5915-21. View

15.

Trachsel H, Ranu R, LONDON I . Regulation of protein synthesis in rabbit reticulocyte lysates: purification and characterization of heme-reversible translational inhibitor. Proc Natl Acad Sci U S A. 1978; 75(8):3654-8. PMC: 392844. DOI: 10.1073/pnas.75.8.3654. View

16.

Farrell P, Hunt T, Jackson R . Analysis of phosphorylation of protein synthesis initiation factor eIF-2 by two-dimensional gel electrophoresis. Eur J Biochem. 1978; 89(2):517-21. DOI: 10.1111/j.1432-1033.1978.tb12556.x. View

17.

Ranu R, LONDON I . Regulation of protein synthesis in rabbit reticulocyte lysates: additional initiation factor required for formation of ternary complex (eIF-2.GTP.Met-tRNAf) and demonstration of inhibitory effect of heme-regulated protein kinase. Proc Natl Acad Sci U S A. 1979; 76(3):1079-83. PMC: 383192. DOI: 10.1073/pnas.76.3.1079. View

18.

de Haro C, Ochoa S . Further studies on the mode of action of the heme-controlled translational inhibitor. Proc Natl Acad Sci U S A. 1979; 76(4):1741-5. PMC: 383466. DOI: 10.1073/pnas.76.4.1741. View

19.

Ernst V, LEVIN D, LONDON I . In situ phosphorylation of the alpha subunit of eukaryotic initiation factor 2 in reticulocyte lysates inhibited by heme deficiency, double-stranded RNA, oxidized glutathione, or the heme-regulated protein kinase. Proc Natl Acad Sci U S A. 1979; 76(5):2118-22. PMC: 383548. DOI: 10.1073/pnas.76.5.2118. View

20.

de Haro C, Ochoa S . Further studies on the mode of action of the heme-controlled translational inhibitor: stimulating protein acts at level of binary complex formation. Proc Natl Acad Sci U S A. 1979; 76(5):2163-4. PMC: 383557. DOI: 10.1073/pnas.76.5.2163. View