1887

Abstract

SUMMARY

We have studied the kinetics of early adenovirus type 5 (Ad5) protein synthesis during lytic infection of KB cells by wild-type () and transformation-defective host-range () mutants. Proteins encoded within four early regions were studied: early region 1A (E1A: 1.5 to 4.5 map units, mu), E1B (4.5 to 11.2 mu), E2A (61.6 to 74.9 mu), and E4 (91.4 to 99.1 mu). Synthesis of E1A products, the first to appear during lytic infection, was detectable within 2 h after injection, reached a peak within the next hour, then declined to very low levels by 7 h post-infection. Synthesis of E2 and E4 proteins began at about 3 h post-infection, was maximal by 6 h and thereafter declined sharply. The E1B 19K and 58K proteins were first detected around 3 h post-infection and, after reaching maximal levels of expression by 8 h, declined to lower levels by 12 h post-infection. Infections with the E1A mutant 3 were characterized by greatly depressed levels of early expression of E1B, E2 and E4 polypeptides but protein synthesis from these regions appeared to recover at late times. The pattern of expression exhibited by the E1B mutant 6 revealed delayed and reduced levels of expression of E1B, E2 and E4 protein synthesis but increased levels of E1A protein synthesis. These results are consistent with the reported role of E1A gene products in the activation of early gene expression and, in addition, suggest that a function encoded in E1B may also influence the expression of Ad5 early genes at early and late times.

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1984-03-01
2024-03-28
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References

  1. Alestrom P., Akusjarvi G., Perricaudet M., Mathews M., Klessig D., Petterson U. 1980; The gene for polypeptide IX of adenovirus type 2 and its unspliced messenger RNA. Cell 19:671–681
    [Google Scholar]
  2. Asselbergs F. A., Mathews M. B., Smart J. E. 1983; Structural characterization of the proteins encoded by adenovirus early region 2A. Journal of Molecular Biology 163:177–207
    [Google Scholar]
  3. Axelrod N. 1978; Phosphoproteins of adenovirus 2. Virology 87:366–383
    [Google Scholar]
  4. Babich A., Nevins J. 1981; The stability of early adenovirus mRNA is controlled by the 72 Kd DNA-binding protein. Cell 26:371–379
    [Google Scholar]
  5. Beltz G. A., Flint S. J. 1979; Inhibition of HeLa cell protein synthesis during adenovirus infection: restriction of cellular messenger RNA sequences to the nucleus. Journal of Molecular Biology 131:353–373
    [Google Scholar]
  6. Berk A. J., Sharp P. A. 1978; Structure of the adenovirus 2 early mRNAs. Cell 14:695–711
    [Google Scholar]
  7. Berk A. J., Lee F., Harrison T., Williams J., Sharp P. A. 1979; Pre-early adenovirus 5 gene product regulates synthesis of early viral messenger RNAs. Cell 17:935–944
    [Google Scholar]
  8. Bos J. L., Polder L. J., Bernards R., Schrier P. I., van Den Elsen P. J., van Der Eb A. J., van Ormondt H. 1981; The 2.2 Kb mRNA of the E1B region of human adenovirus type 5 and 12 directs the synthesis of two major antigens from different AUG triplets. Cell 27:121–131
    [Google Scholar]
  9. Chin W., Maizel J. V. 1977; The polypeptides of adenovirus VIII. The enrichment of E3 (11, 000) in the nuclear matrix fraction. Virology 76:79–89
    [Google Scholar]
  10. Chow L. T., Broker T. R., Lewis J. B. 1979; Complex splicing patterns of RNAs from the early regions of adenovirus 2. Journal of Molecular Biology 141:249–265
    [Google Scholar]
  11. Downey J. D., Rowe D. T., Bacchetti S., Graham F. L., Bayley S. T. 1983; The mapping of a 14K antigen to early region 4 of the human adenovirus 5 genome. Journal of Virology 45:514–523
    [Google Scholar]
  12. Esche H., Mathews M. B., Lewis J. B. 1980; Proteins and messenger RNAs of the transforming region of wt and mutant adenovirus. Journal of Molecular Biology 142:399–417
    [Google Scholar]
  13. Flint S. J. 1977; The topography and transcription of the adenovirus genome. Cell 10:153–166
    [Google Scholar]
  14. Galos R. S., Williams J., Binger M., Flint S. F. 1979; Location of additional early gene sequences in the adenoviral chromosome. Cell 17:945–956
    [Google Scholar]
  15. Galos R. S., Williams J., Shenk T., Jones N. 1980; Physical location of host-range mutations of Ad5: detection and marker-rescue mapping. Virology 104:510–513
    [Google Scholar]
  16. Gingeras T. R., Sciaky D., Geunas R., Bing-Dong J., Yen C. E., Kelly M. M., Bullock P. A., Parsons B. L., O’Neill K. E., Roberts R. I. 1982; Nucleotide sequences from the adenovirus 2 genome. Journal of Biological Chemistry 257:13475–13491
    [Google Scholar]
  17. Ginsberg H. S., Lundholm U., Linne T. 1977; Adenovirus DNA binding proteins in cells infected with wt Ad5 and two DNA-minus, temperature sensitive mutants H5 ts 125 and H5 ts 149. Journal of Virology 23:142–151
    [Google Scholar]
  18. Graham F. L., Smiley I., Russell W. C., Nairn R. 1977; Characteristics of a human cell transformed by DNA from human adenovirus 5. Journal of General Virology 36:59–72
    [Google Scholar]
  19. Graham F. L., Harrison T., Williams I. 1978; Defective transforming capacity of adenovirus 5 host-range mutants. Virology 86:10–21
    [Google Scholar]
  20. Green M., Wold W. S. M., Brackmann K. H., Cartas M. A. 1979; Identification of families of overlapping poly-peptides coded by early ‘transforming’ gene region I of human adenovirus 2. Virology 97:275–286
    [Google Scholar]
  21. Halbert D. N., Spector D. J., Raskas H. J. 1979; In vitro translation products specified by the transforming region of Ad2. Journal of Virology 31:621–629
    [Google Scholar]
  22. Harrison T., Graham F. L., Williams J. 1977; Host-range mutants of adenovirus type 5 defective for growth in HeLa cells. Virology 77:319–329
    [Google Scholar]
  23. Harter M. L., Lewis J. B. 1978; Adenovirus type 2 early proteins synthesized in vitro and in vivo: identification in infected cells of the 38, 000 to 50, 000 molecular weight protein encoded by the left end of the adenovirus type 2 genome. Journal of Virology 26:736–749
    [Google Scholar]
  24. Jeng Y. H., Wold W. S. M., Sugawara K., Gilead Z., Green M. 1977; Adenovirus type 2 coded single stranded nucleic acid binding protein: in vivo phosphorylation and modification. Journal of Virology 22:402–411
    [Google Scholar]
  25. Johansson K., Persson H., Lewis A. M., Petterson U., Tibbits C., Philipson L. 1978; Viral DNA sequences and gene products in hamster cells transformed by Ad2. Journal of Virology 27:628–639
    [Google Scholar]
  26. Jones N., Shenk T. 1979a; An adenovirus type 5 early gene function regulates expression of other early viral genes. Proceedings of the National Academy of Sciences, U. S. A 76:3665–3669
    [Google Scholar]
  27. Jones N., Shenk T. 1979b; Isolation of adenovirus type 5 host-range deletion mutants defective for transformation of rat embryo cells. Cell 17:683–689
    [Google Scholar]
  28. Katze M. G., Persson H., Philipson L. 1981; Control of adenovirus early gene expression: post-transcriptional control mediated by both viral and cellular gene products. Molecular and Cellular Biology 1:807–813
    [Google Scholar]
  29. Killington R. A., Newhook L., Balachandran N., Rawls W. E., Bacchetti S. 1981; Production of hybrid cell lines secreting antibodies to herpes simplex virus type 2. Journal of Virological Methods 2:223–236
    [Google Scholar]
  30. Klessig D. F., Grodzicker T. 1979; Mutations that allow human Ad2 and Ad5 to express late genes in monkey cells map in the viral gene encoding the 72K DNA binding protein. Cell 17:957–966
    [Google Scholar]
  31. Laemmli U.K. 1970; Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature, London 227:680–685
    [Google Scholar]
  32. Lassam N. J., Bayley S. T., Graham F. L. 1978; Synthesis of DNA, late polypeptides, and infectious virus by host-range mutants of Ad5 in non-permissive cells. Virology 87:463–467
    [Google Scholar]
  33. Lassam N. J., Bayley S. T., Graham F. L. 1979; Tumor antigens of human Ad5 in transformed and in cells infected with transformation defective host-range mutants. Cell 18:781–791
    [Google Scholar]
  34. Levine A. J., van Der Vliet P. C., Rosenwirth B., Rabek J., Frankel G., Ensinger M. 1974; Adenovirus infected cell-specific DNA binding proteins. Cold Spring Harbor Symposia on Quantitative Biology 39:559–566
    [Google Scholar]
  35. Levinson A. D., Postel E. H., Levine A. J. 1977; In vivo and in vitro phosphorylation of the adenovirus type 5 single strand specific DNA binding protein. Virology 79:144–159
    [Google Scholar]
  36. Lewis J. B., Mathews M. B. 1980; Control of adenovirus early gene expression: a class of immediate early products. Cell 21:303–313
    [Google Scholar]
  37. Nagata K., Guggenheimer R. A., Enomoto T., Lichy J. H., Hurwitz J. 1982; Adenovirus DNA replication in vitro: identification of a host factor that stimulates synthesis of the preterminal protein-dCMP complex. Proceedings of the National Academy of Sciences, U. S. A 79:6438–6442
    [Google Scholar]
  38. Neuwald P. D., Meyer J., Maizel J. V. Jr, Westphal H. 1977; Early gene expression of adenovirus type 2: R-loop mapping of mRNA and time course of viral DNA, mRNA and protein synthesis. Journal of Virology 21:1019–1030
    [Google Scholar]
  39. Nevins J. R. 1981; Mechanism of activation of early viral transcription by the adenovirus E1A gene product. Cell 26:213–230
    [Google Scholar]
  40. Nevins J. R., Winkler J. J. 1980; Regulation of early adenovirus transcription. A protein product of early region 2 specifically represses region 4 transcription. Proceedings of the National Academy of Sciences, U,. S,. A 77:1893–1897
    [Google Scholar]
  41. Nevins J. R., Ginsberg H. S., Blanchard J. M., Wilson M. C., Darnell J. E. 1979; Regulation of the primary expression of the early adenovirus transcription units. Journal of Virology 32:727–733
    [Google Scholar]
  42. Nicolas J. C., Ingrand D., Sarnow P., Levine A. J. 1982; A mutation in the adenovirus type 5 DNA binding protein that fails to autoregulate the production of the DNA binding protein. Virology 122:481–485
    [Google Scholar]
  43. Perricaudet M., Akusjarvi G., Virtanen A., Petterson U. 1979; Structure of two spliced mRNAs from the transforming region of human subgroup C adenoviruses. Nature, London 281:694–696
    [Google Scholar]
  44. Persson H., Signes C., Philipson L. 1979; Purification and characterization of an early glycoprotein from adenovirus 2 infected cells. Journal of Virology 29:938–948
    [Google Scholar]
  45. Persson J., Iansson M., Philipson L. 1980; Purification and genomic site for an adenovirus type 2 early glycoprotein. Journal of Molecular Biology 136:375–394
    [Google Scholar]
  46. Persson H., Monstein H., Akusjarvi G., Philipson L. 1981; Adenovirus early gene products may control viral mRNA accumulation and translation in vivo. Cell 23:485–496
    [Google Scholar]
  47. Ricciardi R. L., Jones R. L., Cepko C. L., Sharp P. A., Roberts B. E. 1981; Expression of early adenovirus genes requires a viral encoded acidic polypeptide. Proceedings of the National Academy of Sciences, U,. S,. A 78:6121–6125
    [Google Scholar]
  48. Ross S. R., Flint S. J., Levine A. J. 1980a; Identification of the adenovirus early proteins and their genomic map positions. Virology 100:419–432
    [Google Scholar]
  49. Ross S. R., Levine A. J., Galos R. S., Williams J., Shenk T. 1980b; Early viral proteins in HeLa cells infected with adenovirus type 5 host-range mutants. Virology 103:475–492
    [Google Scholar]
  50. Rowe D. T., Graham F. L. 1981; Complementation of adenovirus type 5 host-range mutants by adenovirus type 12 in co-infected HeLa and BHK-21 cells. Journal of Virology 38:191–197
    [Google Scholar]
  51. Rowe D. T., Yee S. P., Otis J., Graham F. L., Branton P. E. 1983a; Characterization of human adenovirus type 5 E1A polypeptides using anti-tumor sera and an antiserum specific for the carboxy terminus. Virology 127:253–271
    [Google Scholar]
  52. Rowe D. T., Graham F. L., Branton P. E. 1983b; Intracellular localization of adenovirus type 5 tumor antigens in productively infected cells. Virology 129:456–468
    [Google Scholar]
  53. Rowe D. T., Branton P. E., Yee S.-P., Bacchetti S., Graham F. L. 1984; Establishment and characterization of hamster cell lines transformed by restriction endonuclease fragments of adenovirus type 5. Journal of Virology (in press)
    [Google Scholar]
  54. Sarnow P., Hearing P., Anderson C. W., Reich N. 1982; Identification and characterization of an immunologically conserved adenovirus early region 11, 000 Mr protein and its association with the nuclear matrix. Journal of Molecular Biology 162:565–587
    [Google Scholar]
  55. Schaffhausen B. S., Silver J. E., Benjamin T. L. 1978; Tumor antigen(s) in cells productively infected by wild-type polyoma virus and mutant N6-18. Proceedings of the National Academy of Sciences, U,. S,. A 75:79–83
    [Google Scholar]
  56. Schrier P. I., van Den Elsen J. J., Hertoghs L., van Der Eb A. J. 1979; Characterization of tumor antigens in cells transformed by fragments of adenovirus type 5 DNA. Virology 99:372–385
    [Google Scholar]
  57. Shaw A. R., Ziff E. B. 1980; Transcripts from the adenovirus 2 major late promoter yield a single early family of 3′ coterminal mRNAs and five late families. Cell 22:905–916
    [Google Scholar]
  58. Shaw A. R., Ziff E. B. 1982; Selective inhibition of adenovirus type 2 early region II and III transcription by an anisomycin block of protein synthesis. Molecular and Cellular Biology 2:789–799
    [Google Scholar]
  59. Smart J. E., Lewis J. B., Mathews M. B., Harter M. L., Anderson C. W. 1981; Adenovirus type 2 early proteins: assignment of the early region 1A proteins synthesized in vivo and in vitro to specific mRNAs. Virology 112:703–713
    [Google Scholar]
  60. Solnick D. 1981; An adenovirus mutant defective in splicing RNA from early region 1A. Nature, London 291:508–510
    [Google Scholar]
  61. Solnick D., Anderson M. A. 1982; Transformation deficient adenovirus mutant defective in expression of region 1A but not region 1B. Journal of Virology 42:106–113
    [Google Scholar]
  62. Spector D. J., McGrogan M., Raskas H. J. 1978; Regulation of the appearance of cytoplasmic RNAs from region 1 of the adenovirus 2 genome. Journal of Molecular Biology 126:395–414
    [Google Scholar]
  63. Stanners C. P., Eliceiri G. L., Green H. 1971; Two types of ribosome in mouse-hamster hybrid cells. Nature New Biology 230:52–54
    [Google Scholar]
  64. Stillman B., Lewis J. B., Chow L. T., Mathews M. B., Smart J. E. 1981; Identification of the gene and mRNA for the adenovirus terminal protein precursor. Cell 23:497–508
    [Google Scholar]
  65. Svensson C., Pettersson U., Akusjarvi G. 1983; Splicing of adenovirus 2 early region 1A mRNAs is non-sequential. Journal of Molecular Biology 165:475–499
    [Google Scholar]
  66. van Der Vliet P. C., Levine A. J. 1973; DNA binding proteins specific for cells infected with adenovirus. Nature New Biology 246:170–174
    [Google Scholar]
  67. van Der Vliet P. C., Levine A. J., Ensinger J. M., Ginsberg H. S. 1975; Thermolabile DNA binding proteins from cells infected with a temperature-sensitive mutant of adenovirus defective in viral DNA synthesis. Journal of Virology 15:348–354
    [Google Scholar]
  68. Williams J. 1973; Oncogenic transformation of hamster embryo cells in vitro by adenovirus type 5. Nature, London 243:162–163
    [Google Scholar]
  69. Wilson M. C., Darnell J. E. 1981; Control of messenger RNA concentration by differential cytoplasmic half-life. Adenovirus messenger RNAs from transcription units 1A and 1B. Journal of Molecular Biology 148:231–251
    [Google Scholar]
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