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. 1988 May;62(5):1606–1616. doi: 10.1128/jvi.62.5.1606-1616.1988

Efficient transcription, not translation, is dependent on adenovirus tripartite leader sequences at late times of infection.

F V Alonso-Caplen 1, M G Katze 1, R M Krug 1
PMCID: PMC253188  PMID: 2833610

Abstract

To determine whether the tripartite leader is required for efficient translation in adenovirus-infected cells at late times of infection, we constructed recombinant adenoviruses containing the influenza virus nucleocapsid protein (NP) gene expressed under the control of the adenovirus major late promoter (MLP). We chose the NP gene because previous results showed that the influenza virus NP mRNA was an extremely effective initiator of translation in cells which were superinfected with influenza virus at late times of adenovirus infection (M. G. Katze, B. M. Detjen, B. Safer, and R. M. Krug, Mol. Cell. Biol. 6:1741-1750, 1986). The NP gene in the adenovirus recombinants was inserted downstream of an MLP that replaced part of the early (E1A) region. The resulting NP mRNAs either lacked any tripartite leader sequences or contained at their 5' ends various portions of the tripartite leader: 33, 172, or all 200 nucleotides of the leader. The relative amounts of the NP protein synthesized by the recombinants were directly proportional to the amounts of the NP mRNA made, indicating that the presence of 5' tripartite leader sequences did not enhance the translation of NP mRNA. In addition, the sizes of the polysomes containing NP mRNA were not increased by the presence of tripartite leader sequences, indicating that the initiation of translation was not enhanced by these sequences. On the other hand, the presence of tripartite leader sequences immediately downstream of the MLP did enhance the transcription of the inserted NP gene, as shown by Northern (RNA) analysis of in vivo NP mRNA levels and by in vitro runoff assays with isolated nuclei. Our results indicate that more than 33 nucleotides of the first leader segment of the tripartite leader are required for optimal transcription from the MLP.

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Selected References

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

  1. Banerjee A. K. 5'-terminal cap structure in eucaryotic messenger ribonucleic acids. Microbiol Rev. 1980 Jun;44(2):175–205. doi: 10.1128/mr.44.2.175-205.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Berget S. M., Moore C., Sharp P. A. Spliced segments at the 5' terminus of adenovirus 2 late mRNA. Proc Natl Acad Sci U S A. 1977 Aug;74(8):3171–3175. doi: 10.1073/pnas.74.8.3171. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Berkner K. L., Sharp P. A. Effect of the tripartite leader on synthesis of a non-viral protein in an adenovirus 5 recombinant. Nucleic Acids Res. 1985 Feb 11;13(3):841–857. doi: 10.1093/nar/13.3.841. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Cervera M., Dreyfuss G., Penman S. Messenger RNA is translated when associated with the cytoskeletal framework in normal and VSV-infected HeLa cells. Cell. 1981 Jan;23(1):113–120. doi: 10.1016/0092-8674(81)90276-2. [DOI] [PubMed] [Google Scholar]
  5. Davis A. R., Kostek B., Mason B. B., Hsiao C. L., Morin J., Dheer S. K., Hung P. P. Expression of hepatitis B surface antigen with a recombinant adenovirus. Proc Natl Acad Sci U S A. 1985 Nov;82(22):7560–7564. doi: 10.1073/pnas.82.22.7560. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Etkind P. R., Krug R. M. Purification of influenza viral complementary RNA: its genetic content and activity in wheat germ cell-free extracts. J Virol. 1975 Dec;16(6):1464–1475. doi: 10.1128/jvi.16.6.1464-1475.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Favaloro J., Treisman R., Kamen R. Transcription maps of polyoma virus-specific RNA: analysis by two-dimensional nuclease S1 gel mapping. Methods Enzymol. 1980;65(1):718–749. doi: 10.1016/s0076-6879(80)65070-8. [DOI] [PubMed] [Google Scholar]
  8. Graham F. L., Smiley J., Russell W. C., Nairn R. Characteristics of a human cell line transformed by DNA from human adenovirus type 5. J Gen Virol. 1977 Jul;36(1):59–74. doi: 10.1099/0022-1317-36-1-59. [DOI] [PubMed] [Google Scholar]
  9. Graham F. L., van der Eb A. J. A new technique for the assay of infectivity of human adenovirus 5 DNA. Virology. 1973 Apr;52(2):456–467. doi: 10.1016/0042-6822(73)90341-3. [DOI] [PubMed] [Google Scholar]
  10. Herz C., Stavnezer E., Krug R., Gurney T., Jr Influenza virus, an RNA virus, synthesizes its messenger RNA in the nucleus of infected cells. Cell. 1981 Nov;26(3 Pt 1):391–400. doi: 10.1016/0092-8674(81)90208-7. [DOI] [PubMed] [Google Scholar]
  11. Jones N., Shenk T. Isolation of adenovirus type 5 host range deletion mutants defective for transformation of rat embryo cells. Cell. 1979 Jul;17(3):683–689. doi: 10.1016/0092-8674(79)90275-7. [DOI] [PubMed] [Google Scholar]
  12. Katze M. G., Chen Y. T., Krug R. M. Nuclear-cytoplasmic transport and VAI RNA-independent translation of influenza viral messenger RNAs in late adenovirus-infected cells. Cell. 1984 Jun;37(2):483–490. doi: 10.1016/0092-8674(84)90378-7. [DOI] [PubMed] [Google Scholar]
  13. Katze M. G., DeCorato D., Krug R. M. Cellular mRNA translation is blocked at both initiation and elongation after infection by influenza virus or adenovirus. J Virol. 1986 Dec;60(3):1027–1039. doi: 10.1128/jvi.60.3.1027-1039.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Katze M. G., Detjen B. M., Safer B., Krug R. M. Translational control by influenza virus: suppression of the kinase that phosphorylates the alpha subunit of initiation factor eIF-2 and selective translation of influenza viral mRNAs. Mol Cell Biol. 1986 May;6(5):1741–1750. doi: 10.1128/mcb.6.5.1741. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Katze M. G., Krug R. M. Metabolism and expression of RNA polymerase II transcripts in influenza virus-infected cells. Mol Cell Biol. 1984 Oct;4(10):2198–2206. doi: 10.1128/mcb.4.10.2198. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Kozak M. How do eucaryotic ribosomes select initiation regions in messenger RNA? Cell. 1978 Dec;15(4):1109–1123. doi: 10.1016/0092-8674(78)90039-9. [DOI] [PubMed] [Google Scholar]
  17. Kozak M. Influences of mRNA secondary structure on initiation by eukaryotic ribosomes. Proc Natl Acad Sci U S A. 1986 May;83(9):2850–2854. doi: 10.1073/pnas.83.9.2850. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Kozak M. Point mutations define a sequence flanking the AUG initiator codon that modulates translation by eukaryotic ribosomes. Cell. 1986 Jan 31;44(2):283–292. doi: 10.1016/0092-8674(86)90762-2. [DOI] [PubMed] [Google Scholar]
  19. Lawrence C. B., Jackson K. J. Translation of adenovirus serotype 2 late messenger RNAs. J Mol Biol. 1982 Dec 5;162(2):317–334. doi: 10.1016/0022-2836(82)90529-0. [DOI] [PubMed] [Google Scholar]
  20. Lawrence W. C., Ginsberg H. S. Intracellular uncoating of type 5 adenovirus deoxyribonucleic acid. J Virol. 1967 Oct;1(5):851–867. doi: 10.1128/jvi.1.5.851-867.1967. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Lenk R., Penman S. The cytoskeletal framework and poliovirus metabolism. Cell. 1979 Feb;16(2):289–301. doi: 10.1016/0092-8674(79)90006-0. [DOI] [PubMed] [Google Scholar]
  22. Lewis E. D., Manley J. L. Control of adenovirus late promoter expression in two human cell lines. Mol Cell Biol. 1985 Sep;5(9):2433–2442. doi: 10.1128/mcb.5.9.2433. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Lodish H. F., Froshauer S. Rates of initiation of protein synthesis by two purified species of vesicular stomatitis virus messenger RNA. J Biol Chem. 1977 Dec 25;252(24):8804–8811. [PubMed] [Google Scholar]
  24. Lodish H. F., Porter M. Translational control of protein synthesis after infection by vesicular stomatitis virus. J Virol. 1980 Dec;36(3):719–733. doi: 10.1128/jvi.36.3.719-733.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Logan J., Shenk T. Adenovirus tripartite leader sequence enhances translation of mRNAs late after infection. Proc Natl Acad Sci U S A. 1984 Jun;81(12):3655–3659. doi: 10.1073/pnas.81.12.3655. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Mansour S. L., Grodzicker T., Tjian R. Downstream sequences affect transcription initiation from the adenovirus major late promoter. Mol Cell Biol. 1986 Jul;6(7):2684–2694. doi: 10.1128/mcb.6.7.2684. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Pelletier J., Sonenberg N. Insertion mutagenesis to increase secondary structure within the 5' noncoding region of a eukaryotic mRNA reduces translational efficiency. Cell. 1985 Mar;40(3):515–526. doi: 10.1016/0092-8674(85)90200-4. [DOI] [PubMed] [Google Scholar]
  28. Rigby P. W., Dieckmann M., Rhodes C., Berg P. Labeling deoxyribonucleic acid to high specific activity in vitro by nick translation with DNA polymerase I. J Mol Biol. 1977 Jun 15;113(1):237–251. doi: 10.1016/0022-2836(77)90052-3. [DOI] [PubMed] [Google Scholar]
  29. Sawadogo M., Roeder R. G. Interaction of a gene-specific transcription factor with the adenovirus major late promoter upstream of the TATA box region. Cell. 1985 Nov;43(1):165–175. doi: 10.1016/0092-8674(85)90021-2. [DOI] [PubMed] [Google Scholar]
  30. White B. A., Bancroft F. C. Cytoplasmic dot hybridization. Simple analysis of relative mRNA levels in multiple small cell or tissue samples. J Biol Chem. 1982 Aug 10;257(15):8569–8572. [PubMed] [Google Scholar]
  31. Zain S., Sambrook J., Roberts R. J., Keller W., Fried M., Dunn A. R. Nucleotide sequence analysis of the leader segments in a cloned copy of adenovirus 2 fiber mRNA. Cell. 1979 Apr;16(4):851–861. doi: 10.1016/0092-8674(79)90100-4. [DOI] [PubMed] [Google Scholar]

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