Skip to main content
NCBI home page
Biochemical Journal logoLink to Biochemical Journal
. 1979 May 15;180(2):379–387. doi: 10.1042/bj1800379

Interaction between membrane functions and protein synthesis in reticulocytes. An elongation-stage inhibitor of protein synthesis extracted from the reticulocyte membrane.

D H Wreschner, M Herzberg
PMCID: PMC1161063  PMID: 486115

Abstract

A component of the reticulocyte cell membrane was found to inhibit protein synthesis severely in a reticulocyte lysate system. An investigation into the mode of action of the membrane inhibitor revealed the following facts. (1) The binding of the tertiary initiation complex (methionyl-tRNAfMet-Initiation Factor 2-GTP) to the 40S ribosomal subunit was unaffected by the membrane inhibitor. (2) The membrane component did not interfere with the binding of the 40S initiation complex to the AUG initiation codon and subsequent attachment of the 60S ribosomal subunit. (3) Elongation of the peptide chain, as assayed by peptidyl-puromycin formation, was markedly affected by the membrane inhibitor. Surprisingly, the membrane component caused a considerable increase in peptidyl-puromycin formation. (4) Reticulocyte ribosomes that had been reisolated by high-speed centrifugation, after preincubation with the membrane component, were found to be highly defective when assayed in a cell-free protein-synthesizing system. These results indicated that an extract of the reticulocyte cell membrane inhibited protein synthesis by interacting with the ribosome and thus interfered with the correct functions of the elongation stage of protein synthesis. The implications of this conclusion are discussed in the light of data showing that a highly purified preparation of the membrane inhibitor also displayed an endonucleolytic activity highly specific for 28S RNA.

Full text

PDF
379

Selected References

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

  1. Civelli O., Vincent A., Buri J. F., Scherrer K. Evidence for a translational inhibitor linked to globin mRNA in untranslated free cytoplasmic messenger ribonucleoprotein complexes. FEBS Lett. 1976 Dec 15;72(1):71–76. doi: 10.1016/0014-5793(76)80815-0. [DOI] [PubMed] [Google Scholar]
  2. Cundliffe E., McQuillen K. Bacterial protein synthesis: the effects of antibiotics. J Mol Biol. 1967 Nov 28;30(1):137–146. doi: 10.1016/0022-2836(67)90249-5. [DOI] [PubMed] [Google Scholar]
  3. Danon D., Goldstein L., Marikovsky Y., Skutelsky E. Use of cationized ferritin as a label of negative charges on cell surfaces. J Ultrastruct Res. 1972 Mar;38(5):500–510. doi: 10.1016/0022-5320(72)90087-1. [DOI] [PubMed] [Google Scholar]
  4. Engelhardt D. L. An inhibitor of protein synthesis in cytoplasmic extracts of density inhibited cells. J Cell Physiol. 1971 Dec;78(3):333–344. doi: 10.1002/jcp.1040780303. [DOI] [PubMed] [Google Scholar]
  5. Fisher L. E., Koch G. Inhibition of protein synthesis by HeLa cell surface peptides. Biochem Biophys Res Commun. 1976 Oct 18;72(4):1229–1236. doi: 10.1016/s0006-291x(76)80147-7. [DOI] [PubMed] [Google Scholar]
  6. Gasko O., Danon D. Deterioration and disappearance of mitochondria during reticulocyte maturation. Exp Cell Res. 1972 Nov;75(1):159–169. doi: 10.1016/0014-4827(72)90532-0. [DOI] [PubMed] [Google Scholar]
  7. Gross M., Rabinovitz M. Partial purification of a translational repressor mediating hemin control of globin synthesis and implication of results on the site of inhibition. Biochem Biophys Res Commun. 1973 Feb 5;50(3):832–838. doi: 10.1016/0006-291x(73)91320-x. [DOI] [PubMed] [Google Scholar]
  8. Herzberg M., Breitbart H., Atlan H. Interactions between membrane functions and protein synthesis in reticulocytes. Effects of valinomycin and dicyclohexyl-18-crown-6. Eur J Biochem. 1974 Jun 1;45(1):161–170. doi: 10.1111/j.1432-1033.1974.tb03540.x. [DOI] [PubMed] [Google Scholar]
  9. Herzberg M., Revel M., Danon D. The influence of ribosomal factors during the maturation of reticulocytes. Eur J Biochem. 1969 Nov;11(1):148–153. doi: 10.1111/j.1432-1033.1969.tb00752.x. [DOI] [PubMed] [Google Scholar]
  10. Hobden A. N., Cundliffe E. The mode of action of alpha sarcin and a novel assay of the puromycin reaction. Biochem J. 1978 Jan 15;170(1):57–61. doi: 10.1042/bj1700057. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Huez G., Marbaix G., Hubert E., Leclercq M., Nudel U., Soreq H., Salomon R., Lebleu B., Revel M., Littauer U. Z. Role of the polyadenylate segment in the translation of globin messenger RNA in Xenopus oocytes. Proc Natl Acad Sci U S A. 1974 Aug;71(8):3143–3146. doi: 10.1073/pnas.71.8.3143. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Inoue M., Okajima K., Ito K., Utsumi K., Seno S. Changes in structural organization of surface membrane during erythrocyte maturation. Biochim Biophys Acta. 1977 Jun 2;467(2):130–136. doi: 10.1016/0005-2736(77)90190-0. [DOI] [PubMed] [Google Scholar]
  13. Koch P. A., Gardner F. H., Carter J. R., Jr Red cell maturation: loss of a reticulocyte-specific membrane protein. Biochem Biophys Res Commun. 1973 Oct 15;54(4):1296–1299. doi: 10.1016/0006-291x(73)91128-5. [DOI] [PubMed] [Google Scholar]
  14. Light N. D., Tanner M. J. Changes in surface-membrane components during the differentation of rabbit erythroid cells. Biochem J. 1977 Jun 15;164(3):565–578. doi: 10.1042/bj1640565. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Noll H. Characterization of macromolecules by constant velocity sedimentation. Nature. 1967 Jul 22;215(5099):360–363. doi: 10.1038/215360a0. [DOI] [PubMed] [Google Scholar]
  16. Schindler D. G., Davies J. E. Specific cleavage of ribosomal RNA caused by alpha sarcin. Nucleic Acids Res. 1977 Apr;4(4):1097–1110. doi: 10.1093/nar/4.4.1097. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Scornik O. A., Hoagland M. B., Pfefferkorn L. C., Bishop E. A. Inhibitors of protein synthesis in rat liver microsome fractions. J Biol Chem. 1967 Jan 10;242(1):131–139. [PubMed] [Google Scholar]
  18. Shattil S. J., Cooper R. A. Maturation of macroreticulocyte membranes in vivo. J Lab Clin Med. 1972 Feb;79(2):215–227. [PubMed] [Google Scholar]
  19. Tavassoli M., Crosby W. H. Fate of the nucleus of the marrow erythroblast. Science. 1973 Mar 2;179(4076):912–913. doi: 10.1126/science.179.4076.912. [DOI] [PubMed] [Google Scholar]
  20. Wreschner D., Herzberg M. Interaction between membrane functions and protein synthesis in reticulocytes. A membrane constituent which inhibits protein synthesis in cell-free system. Eur J Biochem. 1976 May 1;64(2):399–404. doi: 10.1111/j.1432-1033.1976.tb10315.x. [DOI] [PubMed] [Google Scholar]
  21. Wreschner D., Melloul D., Herzberg M. Interaction between membrane functions and protein synthesis in reticulocytes: specific cleavage of 28-S ribosomal RNA by a membrane constituent. Eur J Biochem. 1978 Apr;85(1):233–240. doi: 10.1111/j.1432-1033.1978.tb12231.x. [DOI] [PubMed] [Google Scholar]
  22. Wreschner D., Melloul D., Herzberg M. Specific degradation of ribosomal RNA in rabbit reticulocyte membrane-bound ribosomes. FEBS Lett. 1977 May 1;77(1):83–86. doi: 10.1016/0014-5793(77)80197-x. [DOI] [PubMed] [Google Scholar]

Articles from Biochemical Journal are provided here courtesy of The Biochemical Society

RESOURCES