Skip to main content
PMC home page
Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1978 Feb;75(2):650–654. doi: 10.1073/pnas.75.2.650

Translational control: recognition of the methylated 5' end and an internal sequence in eukaryotic mRNA by the initiation factor that binds methionyl-tRNAfMet.

R Kaempfer, H Rosen, R Israeli
PMCID: PMC411313  PMID: 273228

Abstract

Structural analogs of the methylated 5' end (cap) of eukaryotic mRNA, such as 7-methylguanosine 5'-monophosphate, specifically inhibit both GTP-dependent binding of Met-tRNAfMet and binding of globin mRNA to eukaryotic initiation factor 2 (eIF-2). Addition of purified eIF-2 effectively relieves the cap analog-induced inhibition of globin mRNA translation. The analog competitively inhibits the function of eIF-2 and of mRNA in protein synthesis. Binding to eIF-2 of capped mRNA as well as noncapped mRNA, such as Mengo virus RNA, can be inhibited completely by free cap molecules, but much more cap is needed to inhibit binding of Mengo virus RNA. mRNA, whether or not it is capped, competitively inhibits the binding of Met-tRNAfMet to eIF-2. These results provide compelling evidence that eIF-2 recognizes mRNA. It is shown that binding of mRNA to eIF-2 is primarily at an internal sequence, and secondarily through the cap. A model for the function of eIF-2 is presented that can account for all these properties. This model can provide a molecular basis for the differential translation of mRNA species, whether or not they are capped.

Full text

PDF
650

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. Both G. W., Furuichi Y., Muthukrishnan S., Shatkin A. J. Effect of 5'-terminal structure and base composition on polyribonucleotide binding to ribosomes. J Mol Biol. 1976 Jul 5;104(3):637–658. doi: 10.1016/0022-2836(76)90126-1. [DOI] [PubMed] [Google Scholar]
  3. Both G. W., Furuichi Y., Muthukrishnan S., Shatkin A. J. Ribosome binding to reovirus mRNA in protein synthesis requires 5' terminal 7-methylguanosine. Cell. 1975 Oct;6(2):185–195. doi: 10.1016/0092-8674(75)90009-4. [DOI] [PubMed] [Google Scholar]
  4. Canaani D., Revel M., Groner Y. Translational discrimination of 'capped' and 'non-capped' mRNAS: inhibition of a series of chemical analogs of m7GpppX. FEBS Lett. 1976 May 1;64(2):326–331. doi: 10.1016/0014-5793(76)80321-3. [DOI] [PubMed] [Google Scholar]
  5. Darnbrough C., Legon S., Hunt T., Jackson R. J. Initiation of protein synthesis: evidence for messenger RNA-independent binding of methionyl-transfer RNA to the 40 S ribosomal subunit. J Mol Biol. 1973 May 25;76(3):379–403. doi: 10.1016/0022-2836(73)90511-1. [DOI] [PubMed] [Google Scholar]
  6. 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]
  7. 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]
  8. Jay G., Kaempfer R. Sequence of events in initiation of translation: a role for initiator transfer RNA in the recognition of messenger RNA. Proc Natl Acad Sci U S A. 1974 Aug;71(8):3199–3203. doi: 10.1073/pnas.71.8.3199. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Kaempfer R., Hollender R., Abrams W. R., Israeli R. Specific binding of messenger RNA and methionyl-tRNAfMet by the same initiation factor for eukaryotic protein synthesis. Proc Natl Acad Sci U S A. 1978 Jan;75(1):209–213. doi: 10.1073/pnas.75.1.209. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Kaempfer R., Kaufman J. Inhibition of cellular protein synthesis by double-stranded RNA: inactivation of an initiation factor. Proc Natl Acad Sci U S A. 1973 Apr;70(4):1222–1226. doi: 10.1073/pnas.70.4.1222. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Kaempfer R., Kaufman J. Translational control of hemoglobin synthesis by an initiation factor required for recycling of ribosomes and for their binding to messenger RNA. Proc Natl Acad Sci U S A. 1972 Nov;69(11):3317–3321. doi: 10.1073/pnas.69.11.3317. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Lawrence C., Thach R. E. Encephalomyocarditis virus infection of mouse plasmacytoma cells. I. Inhibition of cellular protein synthesis. J Virol. 1974 Sep;14(3):598–610. doi: 10.1128/jvi.14.3.598-610.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. 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]
  14. 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]
  15. 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]
  16. Nuss D. L., Oppermann H., Koch G. Selective blockage of initiation of host protein synthesis in RNA-virus-infected cells. Proc Natl Acad Sci U S A. 1975 Apr;72(4):1258–1262. doi: 10.1073/pnas.72.4.1258. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. 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]
  18. 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]
  19. 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]
  20. Rottman F., Shatkin A. J., Perry R. P. Sequences containing methylated nucleotides at the 5' termini of messenger RNAs: possible implications for processing. Cell. 1974 Nov;3(3):197–199. doi: 10.1016/0092-8674(74)90131-7. [DOI] [PubMed] [Google Scholar]
  21. Schreier M. H., Staehelin T., Gesteland R. F., Spahr P. F. Translation of bacteriophage R17 and Qbeta RNA in a mammalian cell-free system. J Mol Biol. 1973 Apr 15;75(3):575–578. doi: 10.1016/0022-2836(73)90462-2. [DOI] [PubMed] [Google Scholar]
  22. Schreier M. H., Staehelin T. Initiation of eukaryotic protein synthesis: (Met-tRNA f -40S ribosome) initiation complex catalysed by purified initiation factors in the absence of mRNA. Nat New Biol. 1973 Mar 14;242(115):35–38. doi: 10.1038/newbio242035a0. [DOI] [PubMed] [Google Scholar]
  23. Shafritz D. A., Weinstein J. A., Safer B., Merrick W. C., Weber L. A., Hickey E. D., Baglioni C. Evidence for role of m7G5'-phosphate group in recognition of eukaryotic mRNA by initiation factor IF-M3. Nature. 1976 May 27;261(5558):291–294. doi: 10.1038/261291a0. [DOI] [PubMed] [Google Scholar]
  24. Shatkin A. J. Capping of eucaryotic mRNAs. Cell. 1976 Dec;9(4 Pt 2):645–653. doi: 10.1016/0092-8674(76)90128-8. [DOI] [PubMed] [Google Scholar]
  25. Suzuki H. Effect of 7-methylguanosine-5'-phosphate on rabbit globin synthesis. FEBS Lett. 1976 Dec 31;72(2):309–313. doi: 10.1016/0014-5793(76)80993-3. [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., Reichmann M. E. Pyrophosphate in the 5' terminal position of a viral ribonucleic acid. Science. 1968 Jun 28;160(3835):1452–1454. doi: 10.1126/science.160.3835.1452. [DOI] [PubMed] [Google Scholar]

Articles from Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences

RESOURCES