Skip to main content
PMC home page
Journal of Virology logoLink to Journal of Virology
. 1985 Jul;55(1):206–212. doi: 10.1128/jvi.55.1.206-212.1985

Restricted changes in the adenovirus DNA-binding protein that lead to extended host range or temperature-sensitive phenotypes.

D E Brough, S A Rice, S Sell, D F Klessig
PMCID: PMC254916  PMID: 3925161

Abstract

Human adenovirus fails to multiply efficiently in monkey cells owing to a block to late viral gene expression. Ad2hr400 through Ad2hr403 are a set of host range (hr) mutants which were selected for their ability to readily grow in these cells at 37 degrees C. The mutations responsible for this extended host range have previously been mapped to the 5' portion of the gene encoding the 72-kilodalton DNA-binding protein (DBP). DNA sequence analyses indicate that all four hr mutants contain the same alteration at coding triplet 130, which changes a histidine codon to a tyrosine codon. These results extend those of Anderson et al. (J. Virol. 48:31-39, 1983), which suggested that only this change in the DBP amino acid sequence can expand adenovirus host range to monkey cells. The hr phenotype does not appear to require phosphorylation of this tyrosine residue, since no phosphotyrosine was detected in DBP isolated from Ad2hr400-infected monkey cells. The hr mutants Ad2hr400 through Ad2hr403, however, are cold sensitive for growth in monkey cells. The mutant Ad2ts400, which was derived from Ad2hr400, represents a second class of hr mutants which can grow efficiently in monkey cells at 32.5 degrees C. The cold-resistant hr mutation of Ad2ts400 has previously been mapped to the 5' region of the DBP gene (map units 63.6 through 66). DNA sequence analysis of this region shows that this mutant contains the original hr alteration at coding triplet 130 as well as a second alteration at coding triplet 148, which changes an alanine codon to a valine codon. We suspect that the alterations at amino acids 130 and 148 change the structure of the amino-terminal domain of the DBP, allowing it to better interact with monkey cell components required for late viral gene expression. Ad2ts400 also contains a temperature-sensitive mutation which has previously been mapped to the 3' portion of the DBP gene (map units 61.3 through 63.6). Sequence analysis of this region indicates that the DBP coding triplet 413 has been altered. This change from a serine codon to a proline codon is the same alteration reported in the previously sequenced DBP mutants Ad5ts125 (W. Kruijer et al., Nucleic Acids Res. 9:4439-4457, 1981) and Ad5ts107 (W. Kruijer et al., Virology 124:425-433, 1983). Thus it appears that only a very limited number of changes in either the 5' or the 3' portion of the DBP gene can give rise to the hr or temperature-sensitive phenotypes, respectively.

Full text

PDF
210

Images in this article

208Image
on p.208
209Image
on p.209
209Image
on p.209

Selected References

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

  1. Anderson C. W., Hardy M. M., Dunn J. J., Klessig D. F. Independent, spontaneous mutants of adenovirus type 2-simian virus 40 hybrid Ad2+ND3 that grow efficiently in monkey cells possess indentical mutations in the adenovirus type 2 DNA-binding protein gene. J Virol. 1983 Oct;48(1):31–39. doi: 10.1128/jvi.48.1.31-39.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Anderson C. W. Spontaneous mutants of the adenovirus-simian virus 40 hybrid, Ad2+ND3, that grow efficiently in monkey cells. Virology. 1981 May;111(1):263–269. doi: 10.1016/0042-6822(81)90670-x. [DOI] [PubMed] [Google Scholar]
  3. Anderson K. P., Klessig D. F. Altered mRNA splicing in monkey cells abortively infected with human adenovirus may be responsible for inefficient synthesis of the virion fiber polypeptide. Proc Natl Acad Sci U S A. 1984 Jul;81(13):4023–4027. doi: 10.1073/pnas.81.13.4023. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Anderson K. P., Klessig D. F. Posttranscriptional block to synthesis of a human adenovirus capsid protein in abortively infected monkey cells. J Mol Appl Genet. 1983;2(1):31–43. [PubMed] [Google Scholar]
  5. Anderson K. P., Klessig D. F. Synthesis of human adenovirus early RNA species is similar in productive and abortive infections of monkey and human cells. J Virol. 1982 May;42(2):748–754. doi: 10.1128/jvi.42.2.748-754.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Ariga H., Klein H., Levine A. J., Horwitz M. S. A cleavage product of the adenovirus DNA binding protein is active in DNA replication in vitro. Virology. 1980 Feb;101(1):307–310. doi: 10.1016/0042-6822(80)90510-3. [DOI] [PubMed] [Google Scholar]
  7. Axelrod N. Phosphoproteins of adenovirus 2. Virology. 1978 Jun 15;87(2):366–383. doi: 10.1016/0042-6822(78)90141-1. [DOI] [PubMed] [Google Scholar]
  8. 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]
  9. Baum S. G., Horwitz M. S., Maizel J. V., Jr Studies of the mechanism of enhancement of human adenovirus infection in monkey cells by simian virus 40. J Virol. 1972 Aug;10(2):211–219. doi: 10.1128/jvi.10.2.211-219.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Berk A. J., Sharp P. A. Structure of the adenovirus 2 early mRNAs. Cell. 1978 Jul;14(3):695–711. doi: 10.1016/0092-8674(78)90252-0. [DOI] [PubMed] [Google Scholar]
  11. Carter T. H., Blanton R. A. Autoregulation of adenovirus type 5 early gene expression II. Effect of temperature-sensitive early mutations on virus RNA accumulation. J Virol. 1978 Nov;28(2):450–456. doi: 10.1128/jvi.28.2.450-456.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Carter T. H., Blanton R. A. Possible role of the 72,000 dalton DNA-binding protein in regulation of adenovirus type 5 early gene expression. J Virol. 1978 Feb;25(2):664–674. doi: 10.1128/jvi.25.2.664-674.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Chow L. T., Broker T. R., Lewis J. B. Complex splicing patterns of RNAs from the early regions of adenovirus-2. J Mol Biol. 1979 Oct 25;134(2):265–303. doi: 10.1016/0022-2836(79)90036-6. [DOI] [PubMed] [Google Scholar]
  14. Clinton G. M., Huang A. S. Distribution of phosphoserine, phosphothreonine and phosphotyrosine in proteins of vesicular stomatitis virus. Virology. 1981 Jan 30;108(2):510–514. doi: 10.1016/0042-6822(81)90459-1. [DOI] [PubMed] [Google Scholar]
  15. Crawford L., Leppard K., Lane D., Harlow E. Cellular proteins reactive with monoclonal antibodies directed against simian virus 40 T-antigen. J Virol. 1982 May;42(2):612–620. doi: 10.1128/jvi.42.2.612-620.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. D'Halluin J. C., Cousin C., Niel C., Boulanger P. Characterization of an early temperature-sensitive and cytocidal double mutant of adenovirus type 2. J Gen Virol. 1984 Aug;65(Pt 8):1305–1317. doi: 10.1099/0022-1317-65-8-1305. [DOI] [PubMed] [Google Scholar]
  17. Ensinger M. J., Ginsberg H. S. Selection and preliminary characterization of temperature-sensitive mutants of type 5 adenovirus. J Virol. 1972 Sep;10(3):328–339. doi: 10.1128/jvi.10.3.328-339.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Farber M. S., Baum S. G. Transcription of adenovirus RNA in permissive and nonpermissive infections. J Virol. 1978 Jul;27(1):136–148. doi: 10.1128/jvi.27.1.136-148.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Feldman L. A., Butel J. S., Rapp F. Interaction of a simian papovavirus and adenoviruses. I. Induction of adenovirus tumor antigen during abortive infection of simian cells. J Bacteriol. 1966 Feb;91(2):813–818. doi: 10.1128/jb.91.2.813-818.1966. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Fey G., Lewis J. B., Grodzicker T., Bothwell A. Characterization of a fused protein specified by the adenovirus type 2-simian virus 40 hybrid Ad2+ND1 dp2. J Virol. 1979 Apr;30(1):201–217. doi: 10.1128/jvi.30.1.201-217.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Frackelton A. R., Jr, Ross A. H., Eisen H. N. Characterization and use of monoclonal antibodies for isolation of phosphotyrosyl proteins from retrovirus-transformed cells and growth factor-stimulated cells. Mol Cell Biol. 1983 Aug;3(8):1343–1352. doi: 10.1128/mcb.3.8.1343. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Friedman M. P., Lyons M. J., Ginsberg H. S. Biochemical consequences of type 2 adenovirus and Simian virus 40 double infections of African green monkey kidney cells. J Virol. 1970 May;5(5):586–597. doi: 10.1128/jvi.5.5.586-597.1970. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Ginsberg H. S., Ensinger M. J., Kauffman R. S., Mayer A. J., Lundholm U. Cell transformation: a study of regulation with types 5 and 12 adenovirus temperature-sensitive mutants. Cold Spring Harb Symp Quant Biol. 1975;39(Pt 1):419–426. doi: 10.1101/sqb.1974.039.01.054. [DOI] [PubMed] [Google Scholar]
  24. Grodzicker T., Anderson C., Sharp P. A., Sambrook J. Conditional lethal mutants of adenovirus 2-simian virus 40 hybrids. I. Host range mutants of Ad2+ND1. J Virol. 1974 Jun;13(6):1237–1244. doi: 10.1128/jvi.13.6.1237-1244.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Handa H., Kingston R. E., Sharp P. A. Inhibition of adenovirus early region IV transcription in vitro by a purified viral DNA binding protein. Nature. 1983 Apr 7;302(5908):545–547. doi: 10.1038/302545a0. [DOI] [PubMed] [Google Scholar]
  26. Hashimoto K., Nakajima K., Oda K., Shimojo H. Complementation of translational defect for growth of human adenovirus type 2 in Simian cells by a Simian virus 40-induced factor. J Mol Biol. 1973 Dec 5;81(2):207–223. doi: 10.1016/0022-2836(73)90190-3. [DOI] [PubMed] [Google Scholar]
  27. Henikoff S. Unidirectional digestion with exonuclease III creates targeted breakpoints for DNA sequencing. Gene. 1984 Jun;28(3):351–359. doi: 10.1016/0378-1119(84)90153-7. [DOI] [PubMed] [Google Scholar]
  28. Horwitz M. S. Temperature-sensitive replication of H5ts125 adenovirus DNA in vitro. Proc Natl Acad Sci U S A. 1978 Sep;75(9):4291–4295. doi: 10.1073/pnas.75.9.4291. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Kitchingman G. R., Lai S. P., Westphal H. Loop structures in hybrids of early RNA and the separated strands of adenovirus DNA. Proc Natl Acad Sci U S A. 1977 Oct;74(10):4392–4395. doi: 10.1073/pnas.74.10.4392. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Klein H., Maltzman W., Levine A. J. Structure-function relationships of the adenovirus DNA-binding protein. J Biol Chem. 1979 Nov 10;254(21):11051–11060. [PubMed] [Google Scholar]
  31. Klessig D. F., Anderson C. W. Block to multiplication of adenovirus serotype 2 in monkey cells. J Virol. 1975 Dec;16(6):1650–1668. doi: 10.1128/jvi.16.6.1650-1668.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Klessig D. F., Chow L. T. Incomplete splicing and deficient accumulation of the fiber messenger RNA in monkey cells infected by human adenovirus type 2. J Mol Biol. 1980 May 15;139(2):221–242. doi: 10.1016/0022-2836(80)90306-x. [DOI] [PubMed] [Google Scholar]
  33. Klessig D. F., Grodzicker T. 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. 1979 Aug;17(4):957–966. doi: 10.1016/0092-8674(79)90335-0. [DOI] [PubMed] [Google Scholar]
  34. Klessig D. F., Quinlan M. P. Genetic evidence for separate functional domains on the human adenovirus specified, 72 kd, DNA binding protein. J Mol Appl Genet. 1982;1(4):263–272. [PubMed] [Google Scholar]
  35. Klessig D. F. Two adenovirus mRNAs have a common 5' terminal leader sequence encoded at least 10 kb upstream from their main coding regions. Cell. 1977 Sep;12(1):9–21. doi: 10.1016/0092-8674(77)90181-7. [DOI] [PubMed] [Google Scholar]
  36. Kruijer W., Nicolas J. C., van Schaik F. M., Sussenbach J. S. Structure and function of DNA binding proteins from revertants of adenovirus type 5 mutants with a temperature-sensitive DNA replication. Virology. 1983 Jan 30;124(2):425–433. doi: 10.1016/0042-6822(83)90358-6. [DOI] [PubMed] [Google Scholar]
  37. Kruijer W., Van Schaik F. M., Sussenbach J. S. Nucleotide sequence of the gene encoding adenovirus type 2 DNA binding protein. Nucleic Acids Res. 1982 Aug 11;10(15):4493–4500. doi: 10.1093/nar/10.15.4493. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Kruijer W., van Schaik F. M., Sussenbach J. S. Structure and organization of the gene coding for the DNA binding protein of adenovirus type 5. Nucleic Acids Res. 1981 Sep 25;9(18):4439–4457. doi: 10.1093/nar/9.18.4439. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Lewis A. M., Jr, Levin M. J., Wiese W. H., Crumpacker C. S., Henry P. H. A nondefective (competent) adenovirus-SV40 hybrid isolated from the AD.2-SV40 hybrid population. Proc Natl Acad Sci U S A. 1969 Aug;63(4):1128–1135. doi: 10.1073/pnas.63.4.1128. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Lewis J. B., Anderson C. W., Atkins J. F. Further mapping of late adenovirus genes by cell-free translation of RNA selected by hybridization to specific DNA fragments. Cell. 1977 Sep;12(1):37–44. doi: 10.1016/0092-8674(77)90183-0. [DOI] [PubMed] [Google Scholar]
  41. Maxam A. M., Gilbert W. Sequencing end-labeled DNA with base-specific chemical cleavages. Methods Enzymol. 1980;65(1):499–560. doi: 10.1016/s0076-6879(80)65059-9. [DOI] [PubMed] [Google Scholar]
  42. Messing J. New M13 vectors for cloning. Methods Enzymol. 1983;101:20–78. doi: 10.1016/0076-6879(83)01005-8. [DOI] [PubMed] [Google Scholar]
  43. Nevins J. R., Winkler J. J. Regulation of early adenovirus transcription: a protein product of early region 2 specifically represses region 4 transcription. Proc Natl Acad Sci U S A. 1980 Apr;77(4):1893–1897. doi: 10.1073/pnas.77.4.1893. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. Nicolas J. C., Sarnow P., Girard M., Levine A. J. Host range temperature-conditional mutants in the adenovirus DNA binding protein are defective in the assembly of infectious virus. Virology. 1983 Apr 15;126(1):228–239. doi: 10.1016/0042-6822(83)90474-9. [DOI] [PubMed] [Google Scholar]
  45. RABSON A. S., O'CONOR G. T., BEREZESKY I. K., PAUL F. J. ENHANCEMENT OF ADENOVIRUS GROWTH IN AFRICAN GREEN MONKEY KIDNEY CELL CULTURES BY SV40. Proc Soc Exp Biol Med. 1964 May;116:187–190. doi: 10.3181/00379727-116-29197. [DOI] [PubMed] [Google Scholar]
  46. Reich P. R., Baum S. G., Rose J. A., Rowe W. P., Weissman S. M. Nucleic acid homology studies of adenovirus type 7-SV40 interactions. Proc Natl Acad Sci U S A. 1966 Feb;55(2):336–341. doi: 10.1073/pnas.55.2.336. [DOI] [PMC free article] [PubMed] [Google Scholar]
  47. Rice S. A., Klessig D. F. The function(s) provided by the adenovirus-specified, DNA-binding protein required for viral late gene expression is independent of the role of the protein in viral DNA replication. J Virol. 1984 Jan;49(1):35–49. doi: 10.1128/jvi.49.1.35-49.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  48. Rubenstein F. E., Ginsberg H. S. Transformation characteristics of temperature-sensitive mutants of type 12 adenovirus. Intervirology. 1974;3(3):170–174. doi: 10.1159/000149753. [DOI] [PubMed] [Google Scholar]
  49. Sanger F., Nicklen S., Coulson A. R. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. doi: 10.1073/pnas.74.12.5463. [DOI] [PMC free article] [PubMed] [Google Scholar]
  50. Stillman B. W., White E., Grodzicker T. Independent mutations in Ad2ts111 cause degradation of cellular DNA and defective viral DNA replication. J Virol. 1984 May;50(2):598–605. doi: 10.1128/jvi.50.2.598-605.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  51. Tanaka T., Weisblum B. Construction of a colicin E1-R factor composite plasmid in vitro: means for amplification of deoxyribonucleic acid. J Bacteriol. 1975 Jan;121(1):354–362. doi: 10.1128/jb.121.1.354-362.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  52. Ushiro H., Cohen S. Identification of phosphotyrosine as a product of epidermal growth factor-activated protein kinase in A-431 cell membranes. J Biol Chem. 1980 Sep 25;255(18):8363–8365. [PubMed] [Google Scholar]
  53. Van Der Vliet P. C., Levine A. J., Ensinger M. J., Ginsberg H. S. Thermolabile DNA binding proteins from cells infected with a temperature-sensitive mutant of adenovrius defective in viral DNA synthesis. J Virol. 1975 Feb;15(2):348–354. doi: 10.1128/jvi.15.2.348-354.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  54. Willians J. F., Young C. S., Austin P. E. Genetic analysis of human adenovirus type 5 in permissive and nonpermissive cells. Cold Spring Harb Symp Quant Biol. 1975;39(Pt 1):427–437. doi: 10.1101/sqb.1974.039.01.055. [DOI] [PubMed] [Google Scholar]
  55. van der Vliet P. C., Levine A. J. DNA-binding proteins specific for cells infected by adenovirus. Nat New Biol. 1973 Dec 12;246(154):170–174. doi: 10.1038/newbio246170a0. [DOI] [PubMed] [Google Scholar]

Articles from Journal of Virology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES