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. 1976 Aug;73(8):2752–2756. doi: 10.1073/pnas.73.8.2752

Bacteriophage T3 and T7 early RNAs are translated by eukaryotic 80S ribosomes: active phage T3 coded S-adenosylmethionine cleaving enzyme is synthesized.

C W Anderson, J F Atkins, J J Dunn
PMCID: PMC430727  PMID: 1066688

Abstract

RNA transcribed in vitro from the early region of bacteriophage T3 or T7 was translated by cytoplasmic ribosomes which synthesized protein in cell-free systems prepared from mammalian cells and wheat germ. The proteins synthesized in vitro and their counterparts prepared from infected Escherichia coli comigrate by polyacrylamide gel electrophoresis with sodium dodecyl sulfate and are similarly affected by deletion or amber bacteriophage mutations. Bacteriophage T3 codes for an enzyme that cleaves S-adenosylmethionine and this activity was detected among the products of the mammalian cell-free system. Bacteriophage T3 or T7 RNA, after endoribonuclease III (EC 3.1.4.24) cleavage, gave higher levels of incorporation into phage T3 or T7 polypeptides than when an equivalent amount of the uncleaved RNA was added to the eukaryotic cell-free systems. Methylation of phage T3 or T7 RNAs is apparently not required for translation in either the wheat germ or mammalian cell-free system. The ability of T3 and T7 RNA to be translated in the presence of saturating amounts of natural eukaryotic mRNAs suggests that many prokaryotic genes introduced into mammalian cells might be expressed if they were transcribed in an appropriate form.

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Selected References

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

  1. Anderson C. W., Lewis J. B., Atkins J. F., Gesteland R. F. Cell-free synthesis of adenovirus 2 proteins programmed by fractionated messenger RNA: a comparison of polypeptide products and messenger RNA lengths. Proc Natl Acad Sci U S A. 1974 Jul;71(7):2756–2760. doi: 10.1073/pnas.71.7.2756. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Ashwell M. A., Work T. S. Contrasting effects of cycloheximide on mitochondrial protein synthesis "in vivo" and "in vitro". Biochem Biophys Res Commun. 1968 Sep 30;32(6):1006–1012. doi: 10.1016/0006-291x(68)90129-0. [DOI] [PubMed] [Google Scholar]
  3. Atkins J. F., Lewis J. B., Anderson C. W., Gesteland R. F. Enhanced differential synthesis of proteins in a mammalian cell-free system by addition of polyamines. J Biol Chem. 1975 Jul 25;250(14):5688–5695. [PubMed] [Google Scholar]
  4. Aviv H., Boime I., Loyd B., Leder P. Translation of bacteriophage Q messenger RNA in a murine Krebs 2 ascites tumor cell-free system. Science. 1972 Dec 22;178(4067):1293–1295. doi: 10.1126/science.178.4067.1293. [DOI] [PubMed] [Google Scholar]
  5. 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]
  6. Cory S., Adams J. M. The modified 5'-terminal sequences in messenger RNA of mouse myeloma cells. J Mol Biol. 1975 Dec 25;99(4):519–547. doi: 10.1016/s0022-2836(75)80170-7. [DOI] [PubMed] [Google Scholar]
  7. Dunn J. J., Studier F. W. Effect of RNAase III, cleavage on translation of bacteriophage T7 messenger RNAs. J Mol Biol. 1975 Dec 15;99(3):487–499. doi: 10.1016/s0022-2836(75)80140-9. [DOI] [PubMed] [Google Scholar]
  8. Dunn J. J., Studier F. W. T7 early RNAs are generated by site-specific cleavages. Proc Natl Acad Sci U S A. 1973 May;70(5):1559–1563. doi: 10.1073/pnas.70.5.1559. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Furuichi Y., Morgan M., Muthukrishnan S., Shatkin A. J. Reovirus messenger RNA contains a methylated, blocked 5'-terminal structure: m-7G(5')ppp(5')G-MpCp-. Proc Natl Acad Sci U S A. 1975 Jan;72(1):362–366. doi: 10.1073/pnas.72.1.362. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Gefter M., Hausmann R., Gold M., Hurwitz J. The enzymatic methylation of ribonucleic acid and deoxyribonucleic acid. X. Bacteriophage T3-induced S-adenosylmethionine cleavage. J Biol Chem. 1966 May 10;241(9):1995–2006. [PubMed] [Google Scholar]
  11. Herrlich P., Schweiger M. T3 and T7 bacteriophage deoxyribonucleic acid-directed enzyme synthesis in vitro. J Virol. 1970 Dec;6(6):750–753. doi: 10.1128/jvi.6.6.750-753.1970. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. 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]
  13. Morrison T. G., Lodish H. F. Recognition of protein synthesis initiation signals on bacteriophage ribonucleic acid by mammalian ribosomes. J Biol Chem. 1974 Sep 25;249(18):5860–5866. [PubMed] [Google Scholar]
  14. Niles E. G., Condit R. C. Translational Mapping of Bacteriophage T7 RNAs synthesized in vitro by purified T7 RNA polymerase. J Mol Biol. 1975 Oct 15;98(1):57–67. doi: 10.1016/s0022-2836(75)80101-x. [DOI] [PubMed] [Google Scholar]
  15. Pieczenik G., Model P., Robertson H. D. Sequence and symmetry in ribosome binding sites of bacteriophage f1 RNA. J Mol Biol. 1974 Dec 5;90(2):191–124. doi: 10.1016/0022-2836(74)90368-4. [DOI] [PubMed] [Google Scholar]
  16. 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]
  17. Rosenberg M., Kramer R. A., Steitz J. A. T7 early messenger RNAs are the direct products of ribonuclease III cleavage. J Mol Biol. 1974 Nov 15;89(4):777–782. doi: 10.1016/0022-2836(74)90052-7. [DOI] [PubMed] [Google Scholar]
  18. 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]
  19. Schreier M. H., Staehelin T. Initiation of mammalian protein synthesis: the importance of ribosome and initiation factor quality for the efficiency of in vitro systems. J Mol Biol. 1973 Feb 19;73(3):329–349. doi: 10.1016/0022-2836(73)90346-x. [DOI] [PubMed] [Google Scholar]
  20. Shatkin A. J., Both G. W. Reovirus mRNA: transcription and translation. Cell. 1976 Mar;7(3):305–313. doi: 10.1016/0092-8674(76)90159-8. [DOI] [PubMed] [Google Scholar]
  21. Shine J., Dalgarno L. Determinant of cistron specificity in bacterial ribosomes. Nature. 1975 Mar 6;254(5495):34–38. doi: 10.1038/254034a0. [DOI] [PubMed] [Google Scholar]
  22. Simon M. N., Studier F. W. Physical mapping of the early region of bacteriophage T7 DNA. J Mol Biol. 1973 Sep 15;79(2):249–265. doi: 10.1016/0022-2836(73)90004-1. [DOI] [PubMed] [Google Scholar]
  23. Skare J., Niles E. G., Summers W. C. Localization of the leftmost initiation site for T7 late transcription, in vivo and in vitro. Biochemistry. 1974 Sep 10;13(19):3912–3916. doi: 10.1021/bi00716a015. [DOI] [PubMed] [Google Scholar]
  24. Steitz J. A., Jakes K. How ribosomes select initiator regions in mRNA: base pair formation between the 3' terminus of 16S rRNA and the mRNA during initiation of protein synthesis in Escherichia coli. Proc Natl Acad Sci U S A. 1975 Dec;72(12):4734–4738. doi: 10.1073/pnas.72.12.4734. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. 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]
  26. Studier F. W. Gene 0.3 of bacteriophage T7 acts to overcome the DNA restriction system of the host. J Mol Biol. 1975 May 15;94(2):283–295. doi: 10.1016/0022-2836(75)90083-2. [DOI] [PubMed] [Google Scholar]
  27. Studier F. W., Movva N. R. SAMase gene of bacteriophage T3 is responsible for overcoming host restriction. J Virol. 1976 Jul;19(1):136–145. doi: 10.1128/jvi.19.1.136-145.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]

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