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. 1978 Oct;75(10):4858–4862. doi: 10.1073/pnas.75.10.4858

Protein synthesis in rabbit reticulocytes: characteristics of a ribosomal factor that reverses inhibition of protein synthesis in heme-deficient lysates.

R O Ralston, A Das, A Dasgupta, R Roy, S Palmieri, N K Gupta
PMCID: PMC336220  PMID: 283396

Abstract

A ribosomal salt (0.5 M KCl) wash factor (RF) that reverses inhibition of protein synthesis in heme-deficient reticulocyte lysates has been resolved from the bulk of Met-tRNAfMet-binding factor (EIF-1), Co-EIF-1, and EIF-2 (ternary complex dissociation factor, TDF). The purified RF restores protein synthesis activity of heme-deficient lysates to the level observed in the presence of hemin. No direct correlation exists between amount of EIF-1 activity and ability to reverse inhibition of protein synthesis in heme-deficient lysates. Homogeneous preparations of EIF-1 are completely inactive in reversal of protein synthesis inhibition in heme-deficient lysates. These findings suggest that RF activity is not due to EIF-1 alone but may or may not require EIF-1 as a component of a complex factor.

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Selected References

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  1. Adamson S. D., Herbert E., Godchaux W. Factors affecting the rate of protein synthesis in lysate systems from reticulocytes. Arch Biochem Biophys. 1968 May;125(2):671–683. doi: 10.1016/0003-9861(68)90625-5. [DOI] [PubMed] [Google Scholar]
  2. Anderson W. F., Bosch L., Cohn W. E., Lodish H., Merrick W. C., Weissbach H., Wittmann H. G., Wool I. G. International symposium on protein synthesis. Summary of Fogarty Center-NIH Workshop held in Bethesda, Maryland on 18-20 October, 1976. FEBS Lett. 1977 Apr 1;76(1):1–10. doi: 10.1016/0014-5793(77)80109-9. [DOI] [PubMed] [Google Scholar]
  3. BRUNS G. P., LONDON I. M. THE EFFECT OF HEMIN ON THE SYNTHESIS OF GLOBIN. Biochem Biophys Res Commun. 1965 Jan 18;18:236–242. doi: 10.1016/0006-291x(65)90746-1. [DOI] [PubMed] [Google Scholar]
  4. Balkow K., Mizuno S., Fisher J. M., Rabinovitz M. Hemin control of globin synthesis: effect of a translational repressor on Met-tRNAf binding to the small ribosomal subunit and its relation to the activity and alailability of an initiation factor. Biochim Biophys Acta. 1973 Oct 26;324(3):397–409. doi: 10.1016/0005-2787(73)90284-0. [DOI] [PubMed] [Google Scholar]
  5. Clemens M. J. Functional relationships between a reticulocyte polypeptide-chain-initiation factor (IF-MP) and the translational inhibitor involved in regulation of protein synthesis by haemin. Eur J Biochem. 1976 Jul 1;66(2):413–422. doi: 10.1111/j.1432-1033.1976.tb10531.x. [DOI] [PubMed] [Google Scholar]
  6. Clemens M. J., Henshaw E. C., Rahamimoff H., London I. M. Met-tRNAfMet binding to 40S ribosomal subunits: a site for the regulation of initiation of protein synthesis by hemin. Proc Natl Acad Sci U S A. 1974 Aug;71(8):2946–2950. doi: 10.1073/pnas.71.8.2946. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Das A., Gupta N. K. Protein synthesis in rabbit reticulocytes XX: a supernatant factor (TDI) inhibits ternary complex (Met-tRNAf-EIF-1-GTP) dissociation and Met-tRNAf binding to 40S ribosomes. Biochem Biophys Res Commun. 1977 Oct 24;78(4):1433–1441. doi: 10.1016/0006-291x(77)91453-x. [DOI] [PubMed] [Google Scholar]
  8. Dasgupta A., Majumdar A., George A. D., Gupta N. K. Protein synthesis in rabbit reticulocytes. XV. Isolation of a ribosomal protein factor (CO-EIE-1) which stimulates Met-tRNAfMet binding to EIF-1. Biochem Biophys Res Commun. 1976 Aug 23;71(4):1234–1241. doi: 10.1016/0006-291x(76)90786-5. [DOI] [PubMed] [Google Scholar]
  9. Dasgupta A., Roy R., Palmieri S., Das A., Ralston R., Gupta N. K. Protein synthesis in rabbit reticulocytes XXII+: a heat stable dialyzable factor (EIF-I*) modulates Met-tRNAf binding to EIF-1. Biochem Biophys Res Commun. 1978 Jun 14;82(3):1019–1027. doi: 10.1016/0006-291x(78)90885-9. [DOI] [PubMed] [Google Scholar]
  10. 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]
  11. Grayzel A. I., Hörchner P., London I. M. The stimulation of globin synthesis by heme. Proc Natl Acad Sci U S A. 1966 Mar;55(3):650–655. doi: 10.1073/pnas.55.3.650. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Gross M. Isolation of two initiation factors that can partially reverse the inhibition of protein synthesis due to hemin deficiency or the hemin-controlled translational represssor in rabbit reticulocyte lysates. Arch Biochem Biophys. 1977 Apr 15;180(1):121–129. doi: 10.1016/0003-9861(77)90015-7. [DOI] [PubMed] [Google Scholar]
  13. Gross M., Mendelewski J. Additional evidence that the hemin-controlled translational repressor from rabbit reticulocytes is a protein kinase. Biochem Biophys Res Commun. 1977 Jan 24;74(2):559–569. doi: 10.1016/0006-291x(77)90340-0. [DOI] [PubMed] [Google Scholar]
  14. Gross M. Reversal of the inhibitory action of the hemin-controlled translational repressor by a post-ribosomal supernatant factor from rabbit reticulocyte lysate. Biochem Biophys Res Commun. 1975 Dec 15;67(4):1507–1515. doi: 10.1016/0006-291x(75)90197-7. [DOI] [PubMed] [Google Scholar]
  15. Gupta N. K., Chatterjee B., Chen Y. C., Majumdar A. Protein synthesis in rabbit reticulocytes. A study of Met-tRNA f Met binding factor(s) and Met-tRNA f Met binding to ribosomes and AUG codon. J Biol Chem. 1975 Feb 10;250(3):853–862. [PubMed] [Google Scholar]
  16. Howard G. A., Adamson S. D., Herbert E. Studies on cessation of protein synthesis in a reticulocyte lysate cell-free system. Biochim Biophys Acta. 1970 Jul 16;213(1):237–240. doi: 10.1016/0005-2787(70)90028-6. [DOI] [PubMed] [Google Scholar]
  17. Hunt T., Vanderhoff G., London I. M. Control of globin synthesis: the role of heme. J Mol Biol. 1972 May 28;66(3):471–481. doi: 10.1016/0022-2836(72)90427-5. [DOI] [PubMed] [Google Scholar]
  18. Kramer G., Cimadevilla J. M., 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 Sep;73(9):3078–3082. doi: 10.1073/pnas.73.9.3078. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Kramer G., Henderson A. B., Pinphanichakarn P., Wallis M. H., Hardesty B. Partial reaction of peptide initiation inhibited by phosphorylation of either initiation factor eIF-2 or 40S ribosomal proteins. Proc Natl Acad Sci U S A. 1977 Apr;74(4):1445–1449. doi: 10.1073/pnas.74.4.1445. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. 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]
  21. Legon S., Jackson R. J., Hunt T. Control of protein synthesis in reticulocyte lysates by haemin. Nat New Biol. 1973 Jan 31;241(109):150–152. doi: 10.1038/newbio241150a0. [DOI] [PubMed] [Google Scholar]
  22. Levin D., Ranu R. S., Ernst V., London I. M. 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 Sep;73(9):3112–3116. doi: 10.1073/pnas.73.9.3112. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Majumdar A., Reynolds S., Gupta N. K. Protein synthesis in rabbit reticulocytes. XIII. Lack of mRNA (poly r (A)) binding activity in highly purified EIF-1. Biochem Biophys Res Commun. 1975 Nov 17;67(2):689–695. doi: 10.1016/0006-291x(75)90867-0. [DOI] [PubMed] [Google Scholar]
  24. Majumdar A., Roy R., Das A., Dasgupta A., Gupta N. K. Protein synthesis in rabbit reticulocytes XIX: EIF-2 promotes dissociation of Met-tRNAf-EIF-1-GTP complex and Met-tRNAf binding to 40S ribosomes. Biochem Biophys Res Commun. 1977 Sep 9;78(1):161–169. doi: 10.1016/0006-291x(77)91235-9. [DOI] [PubMed] [Google Scholar]
  25. Rabinovitz M., Freedman M. L., Fisher J. M., Maxwell C. R. Translational control in hemoglobin syntheskis. Cold Spring Harb Symp Quant Biol. 1969;34:567–578. doi: 10.1101/sqb.1969.034.01.064. [DOI] [PubMed] [Google Scholar]
  26. Ranu R. S., Levin D. H., Delaunay J., Ernst V., London I. M. Regulation of protein synthesis in rabbit reticulocyte lysates: characteristics of inhibition of protein synthesis by a translational inhibitor from heme-deficient lysates and its relationship to the initiation factor which binds Met-tRNAf. Proc Natl Acad Sci U S A. 1976 Aug;73(8):2720–2724. doi: 10.1073/pnas.73.8.2720. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Ranu R. S., London I. M., Das A., Dasgupta A., Majumdar A., Ralston R., Roy R., Gupta N. K. Regulation of protein synthesis in rabbit reticulocyte lysates by the heme-regulated protein kinase: inhibition of interaction of Met-tRNAfMet binding factor with another initiation factor in formation of Met-tRNAfMet.40S ribosomal subunit complexes. Proc Natl Acad Sci U S A. 1978 Feb;75(2):745–749. doi: 10.1073/pnas.75.2.745. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Ranu R. S., London I. M. Regulation of protein synthesis in rabbit reticulocyte lysates: purification and initial characterization of the cyclic 3':5'-AMP independent protein kinase of the heme-regulated translational inhibitor. Proc Natl Acad Sci U S A. 1976 Dec;73(12):4349–4353. doi: 10.1073/pnas.73.12.4349. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Trachsel H., Staehelin T. Binding and release of eukaryotic initiation factor eIF-2 and GTP during protein synthesis initiation. Proc Natl Acad Sci U S A. 1978 Jan;75(1):204–208. doi: 10.1073/pnas.75.1.204. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Zucker W. V., Schulman H. M. Stimulation of globin-chain initiation by hemin in the reticulocyte cell-free system. Proc Natl Acad Sci U S A. 1968 Feb;59(2):582–589. doi: 10.1073/pnas.59.2.582. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. de Haro C., Datta A., Ochoa S. Mode of action of the hemin-controlled inhibitor of protein synthesis. Proc Natl Acad Sci U S A. 1978 Jan;75(1):243–247. doi: 10.1073/pnas.75.1.243. [DOI] [PMC free article] [PubMed] [Google Scholar]

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