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. 1977 Jul;74(7):2734–2738. doi: 10.1073/pnas.74.7.2734

Importance of 5'-terminal blocking structure to stabilize mRNA in eukaryotic protein synthesis.

K Shimotohno, Y Kodama, J Hashimoto, K I Miura
PMCID: PMC431268  PMID: 197518

Abstract

The 7-methylguanylic acid residue confronting the 5'-terminal nucleotide of mRNA through two pyrophosphate linkages was completely removed by tobacco pyrophosphatase from mRNAs of cytoplasmic polyhedrosis virus, tobacco mosaic virus (viral RNA), and globin without any scission in the inner part of the RNA chain. Protein synthesis ability in a wheat germ cell-free system was lost after this treatment of all three kinds of mRNA. The initiation complexes for protein synthesis of these three RNAs were not obtained after using tobacco phosphodiesterase-treated mRNA. On incubation of mRNA in a wheat germ extract, the mRNA lacking m7G was quickly degraded from the 5' terminus in an exonucleolytic way, whereas the intact mRNA remained stable. These results show that one of the confronting nucleotide structure's functions is to stabilize the mRNA, to prevent its degradation.

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Selected References

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

  1. Both G. W., Banerjee A. K., Shatkin A. J. Methylation-dependent translation of viral messenger RNAs in vitro. Proc Natl Acad Sci U S A. 1975 Mar;72(3):1189–1193. doi: 10.1073/pnas.72.3.1189. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. FRAENKEL-CONRAT H., SINGER B., TSUGITA A. Purification of viral RNA by means of bentonite. Virology. 1961 May;14:54–58. doi: 10.1016/0042-6822(61)90131-3. [DOI] [PubMed] [Google Scholar]
  3. Filipowicz W., Furuichi Y., Sierra J. M., Muthukrishnan S., Shatkin A. J., Ochoa S. A protein binding the methylated 5'-terminal sequence, m7GpppN, of eukaryotic messenger RNA. Proc Natl Acad Sci U S A. 1976 May;73(5):1559–1563. doi: 10.1073/pnas.73.5.1559. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Furuichi Y., Miura K. A blocked structure at the 5' terminus of mRNA from cytoplasmic polyhedrosis virus. Nature. 1975 Jan 31;253(5490):374–375. doi: 10.1038/253374a0. [DOI] [PubMed] [Google Scholar]
  5. Hewlett M. J., Rose J. K., Baltimore D. 5'-terminal structure of poliovirus polyribosomal RNA is pUp. Proc Natl Acad Sci U S A. 1976 Feb;73(2):327–330. doi: 10.1073/pnas.73.2.327. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. 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]
  7. Hickey E. D., Weber L. A., Baglioni C. Translation of RNA from unfertilised sea urchin eggs does not require methylation and is inhibited by 7-methylguanosine-5'-monophosphate. Nature. 1976 May 6;261(5555):71–72. doi: 10.1038/261071a0. [DOI] [PubMed] [Google Scholar]
  8. Kozak M., Shatkin A. J. Characterization of ribosome-protected fragments from reovirus messenger RNA. J Biol Chem. 1976 Jul 25;251(14):4259–4266. [PubMed] [Google Scholar]
  9. Lee Y. F., Nomoto A., Detjen B. M., Wimmer E. A protein covalently linked to poliovirus genome RNA. Proc Natl Acad Sci U S A. 1977 Jan;74(1):59–63. doi: 10.1073/pnas.74.1.59. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Leung D. W., Gilbert C. W., Smith R. E., Sasavage N. L., Clark J. M., Jr Translation of satellite tobacco necrosis virus ribonucleic acid by an in vitro system from wheat germ. Biochemistry. 1976 Nov 2;15(22):4943–4950. doi: 10.1021/bi00667a030. [DOI] [PubMed] [Google Scholar]
  11. Miura K., Watanabe K., Sugiura M. 5'-Terminal nucleotide sequences of the double-stranded RNA of silkworm cytoplasmic polyhedrosis virus. J Mol Biol. 1974 Jun 15;86(1):31–48. doi: 10.1016/s0022-2836(74)80005-7. [DOI] [PubMed] [Google Scholar]
  12. Muthukrishnan S., Both G. W., Furuichi Y., Shatkin A. J. 5'-Terminal 7-methylguanosine in eukaryotic mRNA is required for translation. Nature. 1975 May 1;255(5503):33–37. doi: 10.1038/255033a0. [DOI] [PubMed] [Google Scholar]
  13. Nomoto A., Lee Y. F., Wimmer E. The 5' end of poliovirus mRNA is not capped with m7G(5')ppp(5')Np. Proc Natl Acad Sci U S A. 1976 Feb;73(2):375–380. doi: 10.1073/pnas.73.2.375. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Ohno T., Okada Y., Shimotohno K., Miura K., Shinshi H. Enzymatic removal of the 5'-terminal methylated blocked structure of tobacco mosaic virus RNA and its effects on infectivity and reconstitution with coat protein. FEBS Lett. 1976 Aug 15;67(2):209–213. doi: 10.1016/0014-5793(76)80368-7. [DOI] [PubMed] [Google Scholar]
  15. Roberts B. E., Paterson B. M. Efficient translation of tobacco mosaic virus RNA and rabbit globin 9S RNA in a cell-free system from commercial wheat germ. Proc Natl Acad Sci U S A. 1973 Aug;70(8):2330–2334. doi: 10.1073/pnas.70.8.2330. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Roberts B. E., Paterson B. M., Sperling R. The cell-free synthesis and assembly of viral specific polypeptides into TMV particles. Virology. 1974 May;59(1):307–313. doi: 10.1016/0042-6822(74)90227-x. [DOI] [PubMed] [Google Scholar]
  17. 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]
  18. Rose J. K., Lodish H. F. Translation in vitro of vesicular stomatitis virus mRNA lacking 5'-terminal 7-methylguanosine. Nature. 1976 Jul 1;262(5563):32–37. doi: 10.1038/262032a0. [DOI] [PubMed] [Google Scholar]
  19. Shimotohno K., Miura K. i. The process of formation of the 5' -terminal modified structure in messenger RNA of cytoplasmic polyhedrosis virus. FEBS Lett. 1976 Apr 15;64(1):204–208. doi: 10.1016/0014-5793(76)80284-0. [DOI] [PubMed] [Google Scholar]
  20. Shimotono K., Miura K. Single-stranded RNA synthesis in vitro by the RNA polymerase associated with cytoplasmic polyhedrosis virus containing double-stranded RNA. J Biochem. 1973 Jul;74(1):117–125. doi: 10.1093/oxfordjournals.jbchem.a130214. [DOI] [PubMed] [Google Scholar]
  21. Shimotono K., Miura K. Transcription of double-stranded RNA in cytoplasmic polyhedrosis virus in vtro. Virology. 1973 May;53(1):283–286. [PubMed] [Google Scholar]
  22. Shinshi H., Miwa M., Kato K., Noguchi M., Matsushima T., Sugimura T. A novel phosphodiesterase from cultured tobacco cells. Biochemistry. 1976 May 18;15(10):2185–2190. doi: 10.1021/bi00655a024. [DOI] [PubMed] [Google Scholar]
  23. Shinshi H., Miwa M., Sugimura T. Enzyme cleaving the 5'-terminal methylated blocked structure of messenger RNA. FEBS Lett. 1976 Jun 1;65(2):254–257. doi: 10.1016/0014-5793(76)80492-9. [DOI] [PubMed] [Google Scholar]
  24. Squires C., Lee F., Bertrand K., Squires C. L., Bronson M. J., Yanofsky C. Nucleotide sequence of the 5' end of tryptophan messenger RNA of Escherichia coli. J Mol Biol. 1976 May 15;103(2):351–381. doi: 10.1016/0022-2836(76)90317-x. [DOI] [PubMed] [Google Scholar]
  25. Wang S., Marcu K. B., Inouye M. Translation of a specific mRNA from Escherichia coli in a eukaryotic cell-free system. Biochem Biophys Res Commun. 1976 Feb 23;68(4):1194–1200. doi: 10.1016/0006-291x(76)90323-5. [DOI] [PubMed] [Google Scholar]
  26. Weber L. A., Feman E. R., Hickey E. D., Williams M. C., Baglioni C. Inhibition of HeLa cell messenger RNA translation by 7-methylguanosine 5'-monophosphate. J Biol Chem. 1976 Sep 25;251(18):5657–5662. [PubMed] [Google Scholar]
  27. Wimmer E., Chang A. Y., Clark J. M., Jr, Reichmann M. E. Sequence studies of satellite tobacco necrosis virus RNA. Isolation and characterization of a 5'-terminal trinucleotide. J Mol Biol. 1968 Nov 28;38(1):59–73. doi: 10.1016/0022-2836(68)90128-9. [DOI] [PubMed] [Google Scholar]

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