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. 1983 Jul;3(7):1212–1221. doi: 10.1128/mcb.3.7.1212

Effect of adenovirus on metabolism of specific host mRNAs: transport control and specific translational discrimination.

A Babich, L T Feldman, J R Nevins, J E Darnell Jr, C Weinberger
PMCID: PMC370112  PMID: 6688459

Abstract

We have studied the adenovirus-induced inhibition of host cell protein synthesis and the effect of infection on the overall metabolism of host cell mRNA during the late phase of adenovirus infection by following the fate of a number of cellular mRNAs complementary to specific cloned DNA segments. At a time in infection when the rate of total cellular protein synthesis is drastically (greater than 90%) reduced, transcription of specific cellular genes is undiminished. However, the transport of newly synthesized cellular mRNA to the cytoplasm is greatly decreased. This decreased appearance of new mRNA in the cytoplasm cannot account for the observed cessation of cell specific protein synthesis, however, since the concentration of several preexisting cellular mRNAs, including the mRNA for actin, remains unchanged throughout the course of infection. The preexisting mRNA is intact, capped, and functional as judged by its ability to direct protein synthesis in vitro in a cap-dependent fashion. The interruption in host translation appears to operate at the level of initiation directly, since we find that fewer ribosomes are associated with a given cellular mRNA after infection than before infection. Furthermore, the in vivo inhibition of cellular protein synthesis does not appear to be the result of competition with viral mRNA, since conditions which prevent the efficient initiation of translation of viral mRNA (infection with a viral mutant) do not result in the recovery of cell translation. Thus, it appears that a late adenovirus gene product directly mediates a shutoff of host protein synthesis.

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Selected References

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

  1. Aleström P., Akusjärvi G., Perricaudet M., Mathews M. B., Klessig D. F., Pettersson U. The gene for polypeptide IX of adenovirus type 2 and its unspliced messenger RNA. Cell. 1980 Mar;19(3):671–681. doi: 10.1016/s0092-8674(80)80044-4. [DOI] [PubMed] [Google Scholar]
  2. Anderson C. W., Baum P. R., Gesteland R. F. Processing of adenovirus 2-induced proteins. J Virol. 1973 Aug;12(2):241–252. doi: 10.1128/jvi.12.2.241-252.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Babich A., Nevins J. R. The stability of early adenovirus mRNA is controlled by the viral 72 kd DNA-binding protein. Cell. 1981 Nov;26(3 Pt 1):371–379. doi: 10.1016/0092-8674(81)90206-3. [DOI] [PubMed] [Google Scholar]
  4. Bello L. J., Ginsberg H. S. Inhibition of host protein synthesis in type 5 adenovirus-infected cells. J Virol. 1967 Oct;1(5):843–850. doi: 10.1128/jvi.1.5.843-850.1967. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Beltz G. A., Flint S. J. Inhibition of HeLa cell protein synthesis during adenovirus infection. Restriction of cellular messenger RNA sequences to the nucleus. J Mol Biol. 1979 Jun 25;131(2):353–373. doi: 10.1016/0022-2836(79)90081-0. [DOI] [PubMed] [Google Scholar]
  6. Brawerman G. Characteristics and significance of the polyadenylate sequence in mammalian messenger RNA. Prog Nucleic Acid Res Mol Biol. 1976;17:117–148. doi: 10.1016/s0079-6603(08)60068-9. [DOI] [PubMed] [Google Scholar]
  7. Darnell J. E., Jr Variety in the level of gene control in eukaryotic cells. Nature. 1982 Jun 3;297(5865):365–371. doi: 10.1038/297365a0. [DOI] [PubMed] [Google Scholar]
  8. Denhardt D. T. A membrane-filter technique for the detection of complementary DNA. Biochem Biophys Res Commun. 1966 Jun 13;23(5):641–646. doi: 10.1016/0006-291x(66)90447-5. [DOI] [PubMed] [Google Scholar]
  9. Ehrenfeld E. Poliovirus-induced inhibition of host-cell protein synthesis. Cell. 1982 Mar;28(3):435–436. doi: 10.1016/0092-8674(82)90195-7. [DOI] [PubMed] [Google Scholar]
  10. Harpold M. M., Wilson M. C., Darnell J. E., Jr Chinese hamster polyadenylated messenger ribonucleic acid: relationship to non-polyadenylated sequences and relative conservation during messenger ribonucleic acid processing. Mol Cell Biol. 1981 Feb;1(2):188–198. doi: 10.1128/mcb.1.2.188. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Hashimoto S. I., Green M. Multiple methylated cap sequences in adenovirus type 2 early mRNA. J Virol. 1976 Nov;20(2):425–435. doi: 10.1128/jvi.20.2.425-435.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Hofer E., Darnell J. E., Jr The primary transcription unit of the mouse beta-major globin gene. Cell. 1981 Feb;23(2):585–593. doi: 10.1016/0092-8674(81)90154-9. [DOI] [PubMed] [Google Scholar]
  13. Jelinek W., Leinwand L. Low molecular weight RNAs hydrogen-bonded to nuclear and cytoplasmic poly(A)-terminated RNA from cultured Chinese hamster ovary cells. Cell. 1978 Sep;15(1):205–214. doi: 10.1016/0092-8674(78)90095-8. [DOI] [PubMed] [Google Scholar]
  14. Kafatos F. C., Jones C. W., Efstratiadis A. Determination of nucleic acid sequence homologies and relative concentrations by a dot hybridization procedure. Nucleic Acids Res. 1979 Nov 24;7(6):1541–1552. doi: 10.1093/nar/7.6.1541. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. 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]
  16. Land H., Grez M., Hauser H., Lindenmaier W., Schütz G. 5'-Terminal sequences of eucaryotic mRNA can be cloned with high efficiency. Nucleic Acids Res. 1981 May 25;9(10):2251–2266. doi: 10.1093/nar/9.10.2251. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Latham H., Darnell J. E. Entrance of mRNA into HeLa cell cytoplasm in puromycin-treated cells. J Mol Biol. 1965 Nov;14(1):13–22. doi: 10.1016/s0022-2836(65)80225-x. [DOI] [PubMed] [Google Scholar]
  18. Leibowitz R., Penman S. Regulation of protein synthesis in HeLa cells. 3. Inhibition during poliovirus infection. J Virol. 1971 Nov;8(5):661–668. doi: 10.1128/jvi.8.5.661-668.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Lindberg U., Persson T., Philipson L. Isolation and characterization of adenovirus messenger ribonucleic acid in productive infection. J Virol. 1972 Nov;10(5):909–919. doi: 10.1128/jvi.10.5.909-919.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Lindberg U., Sundquist B. Isolation of messenger ribonucleoproteins from mammalian cells. J Mol Biol. 1974 Jun 25;86(2):451–468. doi: 10.1016/0022-2836(74)90030-8. [DOI] [PubMed] [Google Scholar]
  21. Moss B., Koczot F. Sequence of methylated nucleotides at the 5'-terminus of adenovirus-specific RNA. J Virol. 1976 Feb;17(2):385–392. doi: 10.1128/jvi.17.2.385-392.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Nevins J. R. Definition and mapping of adenovirus 2 nuclear transcription. Methods Enzymol. 1980;65(1):768–785. doi: 10.1016/s0076-6879(80)65072-1. [DOI] [PubMed] [Google Scholar]
  23. Ovchinnikov L. P., Avanesov A. T., Seriakova T. A., Alzhanova A. T., Radzhabov H. M. A comparison of the RNA-binding proteins with the proteins of polyribosomal messenger ribonucleoproteins in rabbit reticulocytes. Eur J Biochem. 1978 Oct 16;90(3):527–535. doi: 10.1111/j.1432-1033.1978.tb12632.x. [DOI] [PubMed] [Google Scholar]
  24. Parsons J. T., Green M. Biochemical studies on adenovirus multiplication. 18. Resolution of early virus-specific RNA species in Ad 2 infected and transformed cells. Virology. 1971 Jul;45(1):154–162. doi: 10.1016/0042-6822(71)90122-x. [DOI] [PubMed] [Google Scholar]
  25. Philipson L., Pettersson U., Lindberg U., Tibbetts C., Vennström B., Persson T. RNA synthesis and processing in adenovirus-infected cells. Cold Spring Harb Symp Quant Biol. 1975;39(Pt 1):447–456. doi: 10.1101/sqb.1974.039.01.057. [DOI] [PubMed] [Google Scholar]
  26. Philipson L., Wall R., Glickman G., Darnell J. E. Addition of polyadenylate sequences to virus-specific RNA during adenovirus replication. Proc Natl Acad Sci U S A. 1971 Nov;68(11):2806–2809. doi: 10.1073/pnas.68.11.2806. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Price R., Penman S. Transcription of the adenovirus genome by an -amanitine-sensitive ribonucleic acid polymerase in HeLa cells. J Virol. 1972 Apr;9(4):621–626. doi: 10.1128/jvi.9.4.621-626.1972. [DOI] [PMC free article] [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. 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]
  30. 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]
  31. Skup D., Zarbl H., Millward S. Regulation of translation in L-cells infected with reovirus. J Mol Biol. 1981 Sep 5;151(1):35–55. doi: 10.1016/0022-2836(81)90220-5. [DOI] [PubMed] [Google Scholar]
  32. Soeiro R., Darnell J. E. Competition hybridization by "pre-saturation" of HeLa cell DNA. J Mol Biol. 1969 Sep 28;44(3):551–562. doi: 10.1016/0022-2836(69)90379-9. [DOI] [PubMed] [Google Scholar]
  33. Sommer S., Salditt-Georgieff M., Bachenheimer S., Darnell J. E., Furuichi Y., Morgan M., Shatkin A. J. The methylation of adenovirus-specific nuclear and cytoplasmic RNA. Nucleic Acids Res. 1976 Mar;3(3):749–765. doi: 10.1093/nar/3.3.749. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Thimmappaya B., Weinberger C., Schneider R. J., Shenk T. Adenovirus VAI RNA is required for efficient translation of viral mRNAs at late times after infection. Cell. 1982 Dec;31(3 Pt 2):543–551. doi: 10.1016/0092-8674(82)90310-5. [DOI] [PubMed] [Google Scholar]
  35. Thomas P. S. Hybridization of denatured RNA and small DNA fragments transferred to nitrocellulose. Proc Natl Acad Sci U S A. 1980 Sep;77(9):5201–5205. doi: 10.1073/pnas.77.9.5201. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Trachsel H., Sonenberg N., Shatkin A. J., Rose J. K., Leong K., Bergmann J. E., Gordon J., Baltimore D. Purification of a factor that restores translation of vesicular stomatitis virus mRNA in extracts from poliovirus-infected HeLa cells. Proc Natl Acad Sci U S A. 1980 Feb;77(2):770–774. doi: 10.1073/pnas.77.2.770. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Villa-Komaroff L., Efstratiadis A., Broome S., Lomedico P., Tizard R., Naber S. P., Chick W. L., Gilbert W. A bacterial clone synthesizing proinsulin. Proc Natl Acad Sci U S A. 1978 Aug;75(8):3727–3731. doi: 10.1073/pnas.75.8.3727. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Weber J., Jelinek W., Darnell J. E., Jr The definition of a large viral transcription unit late in Ad2 infection of HeLa cells: mapping of nascent RNA molecules labeled in isolated nuclei. Cell. 1977 Apr;10(4):611–616. doi: 10.1016/0092-8674(77)90093-9. [DOI] [PubMed] [Google Scholar]
  39. Willems M., Penman S. The mechanism of host cell protein synthesis inhibition by poliovirus. Virology. 1966 Nov;30(3):355–367. doi: 10.1016/0042-6822(66)90114-0. [DOI] [PubMed] [Google Scholar]
  40. Wilson M. C., Sawicki S. G., White P. A., Darnell J. E., Jr A correlation between the rate of poly(A) shortening and half-life of messenger RNA in adenovirus transformed cells. J Mol Biol. 1978 Nov 25;126(1):23–36. doi: 10.1016/0022-2836(78)90277-2. [DOI] [PubMed] [Google Scholar]
  41. Wold W. S., Green M., Munns T. W. Methylation of late adenovirus 2 nuclear and messenger RNA. Biochem Biophys Res Commun. 1976 Feb 9;68(3):643–649. doi: 10.1016/0006-291x(76)91194-3. [DOI] [PubMed] [Google Scholar]
  42. Ziff E. B. Transcription and RNA processing by the DNA tumour viruses. Nature. 1980 Oct 9;287(5782):491–499. doi: 10.1038/287491a0. [DOI] [PubMed] [Google Scholar]

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