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. 1983 Sep 24;11(18):6353–6368. doi: 10.1093/nar/11.18.6353

Possible involvement of poly(A) in protein synthesis.

A Jacobson, M Favreau
PMCID: PMC326378  PMID: 6137807

Abstract

The experiments of this paper have re-evaluated the possibility that poly(A) is involved in protein synthesis by testing whether purified poly(A) might competitively inhibit in vitro protein synthesis in rabbit reticulocyte extracts. We have found that poly(A) inhibits the rate of translation of many different poly(A)+ mRNAs and that comparable inhibition is not observed with other ribopolymers. Inhibition by poly(A) preferentially affects the translation of adenylated mRNAs and can be overcome by increased mRNA concentrations or by translating mRNPs instead of mRNA. The extent of inhibition is dependent on the size of the competitor poly(A) as well as on the translation activity which a lysate has for poly(A)+ RNA. In light of our results and numerous experiments in the literature, we propose that poly(A) has a function in protein synthesis and that any role in the determination of mRNA stability is indirect.

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Selected References

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

  1. Adams D. S., Jeffery W. R. Poly(adenylic acid) degradation by two distinct processes in the cytoplasmic RNA of Physarum polycephalum. Biochemistry. 1978 Oct 17;17(21):4519–4524. doi: 10.1021/bi00614a025. [DOI] [PubMed] [Google Scholar]
  2. Armstrong J. A., Edmonds M., Nakazato H., Phillips B. A., Vaughn M. H. Polyadenylic acid sequences in the virion RNA of poliovirus and Eastern Equine Encephalitis virus. Science. 1972 May 5;176(4034):526–528. doi: 10.1126/science.176.4034.526. [DOI] [PubMed] [Google Scholar]
  3. Baer B. W., Kornberg R. D. Repeating structure of cytoplasmic poly(A)-ribonucleoprotein. Proc Natl Acad Sci U S A. 1980 Apr;77(4):1890–1892. doi: 10.1073/pnas.77.4.1890. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Blobel G. A protein of molecular weight 78,000 bound to the polyadenylate region of eukaryotic messenger RNAs. Proc Natl Acad Sci U S A. 1973 Mar;70(3):924–928. doi: 10.1073/pnas.70.3.924. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Brawerman G., Diez J. Metabolism of the polyadenylate sequence of nuclear RNA and messenger RNA in mammalian cells. Cell. 1975 Jul;5(3):271–280. doi: 10.1016/0092-8674(75)90102-6. [DOI] [PubMed] [Google Scholar]
  6. Brawerman G. The Role of the poly(A) sequence in mammalian messenger RNA. CRC Crit Rev Biochem. 1981;10(1):1–38. doi: 10.3109/10409238109114634. [DOI] [PubMed] [Google Scholar]
  7. Colot H. V., Rosbash M. Behavior of individual maternal pA+ RNAs during embryogenesis of Xenopus laevis. Dev Biol. 1982 Nov;94(1):79–86. doi: 10.1016/0012-1606(82)90070-7. [DOI] [PubMed] [Google Scholar]
  8. Deshpande A. K., Chatterjee B., Roy A. K. Translation and stability of rat liver messenger RNA for alpha 2 mu-globulin in Xenopus oocyte. The role of terminal poly(A). J Biol Chem. 1979 Sep 25;254(18):8937–8942. [PubMed] [Google Scholar]
  9. Doel M. T., Carey N. H. The translational capacity of deadenylated ovalbumin messenger RNA. Cell. 1976 May;8(1):51–58. doi: 10.1016/0092-8674(76)90184-7. [DOI] [PubMed] [Google Scholar]
  10. Ehrenfeld E., Summers D. F. Adenylate-rich sequences in vesicular stomatitis virus messenger ribonucleic acid. J Virol. 1972 Oct;10(4):683–688. doi: 10.1128/jvi.10.4.683-688.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Fernandez Muñoz R., Darnell J. E. Poly(A) in mRNA does not contribute to secondary structure necessary for protein synthesis. Cell. 1974 Aug;2(4):247–252. doi: 10.1016/0092-8674(74)90018-x. [DOI] [PubMed] [Google Scholar]
  12. Geoghegan T. E., Sonenshein G. E., Brawerman G. Characteristics and polyadenylate content of the actin messenger RNA of mouse sarcoma-180 ascites cells. Biochemistry. 1978 Oct 3;17(20):4200–4207. doi: 10.1021/bi00613a014. [DOI] [PubMed] [Google Scholar]
  13. Greenberg J. R. Proteins crosslinked to messenger RNA by irradiating polyribosomes with ultraviolet light. Nucleic Acids Res. 1980 Dec 11;8(23):5685–5701. doi: 10.1093/nar/8.23.5685. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Harris B., Dure L., 3rd Developmental regulation in cotton seed germination: polyadenylation of stored messenger RNA. Biochemistry. 1978 Aug 8;17(16):3250–3256. doi: 10.1021/bi00609a012. [DOI] [PubMed] [Google Scholar]
  15. Hickey E. D., Weber L. A., Baglioni C. Inhibition of initiation of protein synthesis by 7-methylguanosine-5'-monophosphate. Proc Natl Acad Sci U S A. 1976 Jan;73(1):19–23. doi: 10.1073/pnas.73.1.19. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Huez G., Marbaix G., Burny A., Hubert E., Leclercq M., Cleuter Y., Chantrenne H., Soreq H., Littauer U. Z. Degradation of deadenylated rabbit alpha-globin mRNA in Xenopus oocytes is associated with its translation. Nature. 1977 Mar 31;266(5601):473–474. doi: 10.1038/266473a0. [DOI] [PubMed] [Google Scholar]
  17. Huez G., Marbaix G., Gallwitz D., Weinberg E., Devos R., Hubert E., Cleuter Y. Functional stabilisation of HeLa cell histone messenger RNAs injected into Xenopus oocytes by 3'-OH polyadenylation. Nature. 1978 Feb 9;271(5645):572–573. doi: 10.1038/271572a0. [DOI] [PubMed] [Google Scholar]
  18. 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]
  19. Jeffery W. R., Brawerman G. Characterization of the steady-state population of messenger RNA and its poly(adenylic acid) segment in mammalian cells. Biochemistry. 1974 Oct 22;13(22):4633–4637. doi: 10.1021/bi00719a026. [DOI] [PubMed] [Google Scholar]
  20. Karpetsky T. P., Boguski M. S., Levy C. C. Structures, properties, and possible biologic functions of polyadenylic acid. Subcell Biochem. 1979;6:1–116. doi: 10.1007/978-1-4615-7945-8_1. [DOI] [PubMed] [Google Scholar]
  21. Kish V. M., Pederson T. Poly (A)-rich ribonucleoprotein complexes from HeLa cell messenger RNA. J Biol Chem. 1976 Oct 10;251(19):5888–5894. [PubMed] [Google Scholar]
  22. Kumar A., Pederson T. Comparison of proteins bound to heterogeneous nuclear RNA and messenger RNA in HeLa cells. J Mol Biol. 1975 Aug 15;96(3):353–365. doi: 10.1016/0022-2836(75)90165-5. [DOI] [PubMed] [Google Scholar]
  23. 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]
  24. Laskey R. A., Mills A. D. Quantitative film detection of 3H and 14C in polyacrylamide gels by fluorography. Eur J Biochem. 1975 Aug 15;56(2):335–341. doi: 10.1111/j.1432-1033.1975.tb02238.x. [DOI] [PubMed] [Google Scholar]
  25. Lehrach H., Diamond D., Wozney J. M., Boedtker H. RNA molecular weight determinations by gel electrophoresis under denaturing conditions, a critical reexamination. Biochemistry. 1977 Oct 18;16(21):4743–4751. doi: 10.1021/bi00640a033. [DOI] [PubMed] [Google Scholar]
  26. Lodish H. F. Model for the regulation of mRNA translation applied to haemoglobin synthesis. Nature. 1974 Oct 4;251(5474):385–388. doi: 10.1038/251385a0. [DOI] [PubMed] [Google Scholar]
  27. Lodish H. F., Nathan D. G. Regulation of hemoglobin synthesis. Preferential inhibition of and globin synthesis. J Biol Chem. 1972 Dec 10;247(23):7822–7829. [PubMed] [Google Scholar]
  28. Maizels N. Dictyostelium 17S, 25S, and 5S rDNAs lie within a 38,000 base pair repeated unit. Cell. 1976 Nov;9(3):431–438. doi: 10.1016/0092-8674(76)90088-x. [DOI] [PubMed] [Google Scholar]
  29. Marbaix G., Huez G., Burny A., Cleuter Y., Hubert E., Leclercq M., Chantrenne H., Soreq H., Nudel U., Littauer U. Z. Absence of polyadenylate segment in globin messenger RNA accelerates its degradation in Xenopus oocytes. Proc Natl Acad Sci U S A. 1975 Aug;72(8):3065–3067. doi: 10.1073/pnas.72.8.3065. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Nemer M., Dubroff L. M., Graham M. Properties of sea urchin embryo messenger RNA containing and lacking poly(A). Cell. 1975 Oct;6(2):171–178. doi: 10.1016/0092-8674(75)90007-0. [DOI] [PubMed] [Google Scholar]
  31. Nudel U., Soreq H., Littauer U. Z. Globin mRNA species containing poly(A) segments of different lengths. Their functional stability in Xenopus oocytes. Eur J Biochem. 1976 Apr 15;64(1):115–121. doi: 10.1111/j.1432-1033.1976.tb10279.x. [DOI] [PubMed] [Google Scholar]
  32. Palatnik C. M., Storti R. V., Capone A. K., Jacobson A. Messenger RNA stability in Dictyostelium discoideum: does poly(A) have a regulatory role? J Mol Biol. 1980 Aug 5;141(2):99–118. doi: 10.1016/0022-2836(80)90379-4. [DOI] [PubMed] [Google Scholar]
  33. Palatnik C. M., Storti R. V., Jacobson A. Fractionation and functional analysis of newly synthesized and decaying messenger RNAs from vegetative cells of Dictyostelium discoideum. J Mol Biol. 1979 Mar 5;128(3):371–395. doi: 10.1016/0022-2836(79)90093-7. [DOI] [PubMed] [Google Scholar]
  34. Pelham H. R., Jackson R. J. An efficient mRNA-dependent translation system from reticulocyte lysates. Eur J Biochem. 1976 Aug 1;67(1):247–256. doi: 10.1111/j.1432-1033.1976.tb10656.x. [DOI] [PubMed] [Google Scholar]
  35. Preston C. M., Szilagyi J. F. Cell-free translation of RNA synthesized in vitro by a transcribing nucleoprotein complex prepared from purified vesicular stomatitis virus. J Virol. 1977 Mar;21(3):1002–1009. doi: 10.1128/jvi.21.3.1002-1009.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Pridgen C., Kingsbury D. W. Adenylate-rich sequences in Sendai virus transcripts from infected cells. J Virol. 1972 Aug;10(2):314–317. doi: 10.1128/jvi.10.2.314-317.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Roman R., Brooker J. D., Seal S. N., Marcus A. Inhibition of the transition of a 40 S ribosome-Met-tRNA-i-Met complex to an 80 S ribosome-Met-tRNA-i-Met- complex by 7-Methylguanosine-5'-phosphate. Nature. 1976 Mar 25;260(5549):359–360. doi: 10.1038/260359a0. [DOI] [PubMed] [Google Scholar]
  38. Rose J. K., Lodish H. F., Brock M. L. Giant heterogeneous polyadenylic acid on vesicular stomatitis virus mRNA synthesized in vitro in the presence of S-adenosylhomocysteine. J Virol. 1977 Feb;21(2):683–693. doi: 10.1128/jvi.21.2.683-693.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Rosenthal E. T., Tansey T. R., Ruderman J. V. Sequence-specific adenylations and deadenylations accompany changes in the translation of maternal messenger RNA after fertilization of Spisula oocytes. J Mol Biol. 1983 May 25;166(3):309–327. doi: 10.1016/s0022-2836(83)80087-4. [DOI] [PubMed] [Google Scholar]
  40. Schwartz H., Darnell J. E. The association of protein with the polyadenylic acid of HeLa cell messenger RNA: evidence for a "transport" role of a 75,000 molecular weight polypeptide. J Mol Biol. 1976 Jul 15;104(4):833–851. doi: 10.1016/0022-2836(76)90185-6. [DOI] [PubMed] [Google Scholar]
  41. Sehgal P. B., Soreq H., Tamm I. Does 3'-terminal poly(A) stabilize human fibroblast interferon mRNA in oocytes of Xenopus laevis? Proc Natl Acad Sci U S A. 1978 Oct;75(10):5030–5033. doi: 10.1073/pnas.75.10.5030. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Setyono B., Greenberg J. R. Proteins associated with poly(A) and other regions of mRNA and hnRNA molecules as investigated by crosslinking. Cell. 1981 Jun;24(3):775–783. doi: 10.1016/0092-8674(81)90103-3. [DOI] [PubMed] [Google Scholar]
  43. Sheiness D., Darnell J. E. Polyadenylic acid segment in mRNA becomes shorter with age. Nat New Biol. 1973 Feb 28;241(113):265–268. doi: 10.1038/newbio241265a0. [DOI] [PubMed] [Google Scholar]
  44. Sippel A. E., Stavrianopoulos J. G., Schutz G., Feigelson P. Translational properties of rabbit globin mRNA after specific removal of poly(A) with ribonuclease H. Proc Natl Acad Sci U S A. 1974 Nov;71(11):4635–4639. doi: 10.1073/pnas.71.11.4635. [DOI] [PMC free article] [PubMed] [Google Scholar]
  45. Slater I., Gillespie D., Slater D. W. Cytoplasmic adenylylation and processing of maternal RNA. Proc Natl Acad Sci U S A. 1973 Feb;70(2):406–411. doi: 10.1073/pnas.70.2.406. [DOI] [PMC free article] [PubMed] [Google Scholar]
  46. Sonenshein G. E., Brawerman G. Differential translation of mouse myeloma messenger RNAs in a wheat germ cell-free system. Biochemistry. 1976 Dec 14;15(25):5501–5506. doi: 10.1021/bi00670a013. [DOI] [PubMed] [Google Scholar]
  47. Sonenshein G. E., Brawerman G. Differential translation of rat liver albumin messenger RNA in a wheat germ cell-free system. Biochemistry. 1977 Dec 13;16(25):5445–5448. doi: 10.1021/bi00644a007. [DOI] [PubMed] [Google Scholar]
  48. Soreq H., Nudel U., Salomon R., Revel M., Littauer U. Z. In vitro translation of polyadenylic acid-free rabbit globin messenger RNA. J Mol Biol. 1974 Sep 5;88(1):233–245. doi: 10.1016/0022-2836(74)90307-6. [DOI] [PubMed] [Google Scholar]
  49. Soria M., Huang A. S. Association of polyadenylic acid with messenger RNA of vesicular stomatitis virus. J Mol Biol. 1973 Jul 5;77(3):449–455. doi: 10.1016/0022-2836(73)90450-6. [DOI] [PubMed] [Google Scholar]
  50. Spector D. H., Villa-Komaroff L., Baltimore D. Studies on the function of polyadenylic acid on poliovirus RNA. Cell. 1975 Sep;6(1):41–44. doi: 10.1016/0092-8674(75)90071-9. [DOI] [PubMed] [Google Scholar]
  51. Storti R. V., Scott M. P., Rich A., Pardue M. L. Translational control of protein synthesis in response to heat shock in D. melanogaster cells. Cell. 1980 Dec;22(3):825–834. doi: 10.1016/0092-8674(80)90559-0. [DOI] [PubMed] [Google Scholar]
  52. Studier F. W. Analysis of bacteriophage T7 early RNAs and proteins on slab gels. J Mol Biol. 1973 Sep 15;79(2):237–248. doi: 10.1016/0022-2836(73)90003-x. [DOI] [PubMed] [Google Scholar]
  53. Weiss S. R., Bratt M. A. Polyadenylate sequences on Newcastle disease virus mRNA synthesized in vivo and in vitro. J Virol. 1974 Jun;13(6):1220–1230. doi: 10.1128/jvi.13.6.1220-1230.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  54. Williamson R., Crossley J., Humphries S. Translation of mouse globin messenger ribonucleic acid from which the poly(adenylic acid) sequence has been removed. Biochemistry. 1974 Feb 12;13(4):703–707. doi: 10.1021/bi00701a011. [DOI] [PubMed] [Google Scholar]
  55. Zeevi M., Nevins J. R., Darnell J. E., Jr Nuclear RNA is spliced in the absence of poly(A) addition. Cell. 1981 Oct;26(1 Pt 1):39–46. doi: 10.1016/0092-8674(81)90031-3. [DOI] [PubMed] [Google Scholar]

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