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. 1981 Sep 25;9(18):4439–4457. doi: 10.1093/nar/9.18.4439

Structure and organization of the gene coding for the DNA binding protein of adenovirus type 5.

W Kruijer, F M van Schaik, J S Sussenbach
PMCID: PMC327449  PMID: 6117824

Abstract

We have determined the nucleotide sequence of a region of adenovirus type 5 (Ad5) DNA located between map positions 61.7 and 71.4, which covers the gene form the 72 kD DNA binding protein (DBP) and the sequence encoding the amino-terminal part of the 100 kD protein. Sequence analysis of cDNA copies of DBP mRNA revealed the existence of two abundant species of spliced mRNA molecules. One species consists of two short leader sequences from positions 75.2 (67 and 68 nucleotides long) and 68.8 (77 nucleotides long), respectively, and the main body of the RNA molecules. The other species contains only the leader sequence from position 75.2 and the main body. The amino acid sequence of DBP is encoded entirely by a long open reading frame of 1587 nucleotides in the main body of DBP mRNA. From the nucleotide sequence of the DBP gene it can be derived that DBP contains 529 amino acid residues and has an actual molecular weight of 59,049 daltons. The sites of mutation in the mutants H5hr404 and H5ts125 were determined at the nucleotide level. Single nucleotide alterations were detected in H5hr404 and H5ts125 in the sequences corresponding to the amino-terminal part and the carboxy-terminal part of DBP, respectively. The implications of these mutations are discussed.

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Selected References

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

  1. 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]
  2. Baker C. C., Herisse J., Courtois G., Galibert F., Ziff E. Messenger RNA for the Ad2 DNA binding protein: DNA sequences encoding the first leader and heterogenity at the mRNA 5' end. Cell. 1979 Oct;18(2):569–580. doi: 10.1016/0092-8674(79)90073-4. [DOI] [PubMed] [Google Scholar]
  3. 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]
  4. Brawerman G., Mendecki J., Lee S. Y. A procedure for the isolation of mammalian messenger ribonucleic acid. Biochemistry. 1972 Feb 15;11(4):637–641. doi: 10.1021/bi00754a027. [DOI] [PubMed] [Google Scholar]
  5. 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]
  6. 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]
  7. Chow L. T., Broker T. R. The spliced structures of adenovirus 2 fiber message and the other late mRNAs. Cell. 1978 Oct;15(2):497–510. doi: 10.1016/0092-8674(78)90019-3. [DOI] [PubMed] [Google Scholar]
  8. 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]
  9. Fowlkes D. M., Lord S. T., Linné T., Pettersson U., Philipson L. Interaction between the adenovirus DNA-binding protein and double-stranded DNA. J Mol Biol. 1979 Aug 5;132(2):163–180. doi: 10.1016/0022-2836(79)90389-9. [DOI] [PubMed] [Google Scholar]
  10. Frost E., Williams J. Mapping temperature-sensitive and host-range mutations of adenovirus type 5 by marker rescue. Virology. 1978 Nov;91(1):39–50. doi: 10.1016/0042-6822(78)90353-7. [DOI] [PubMed] [Google Scholar]
  11. Galibert F., Hérissé J., Courtois G. Nucleotide sequence of the EcoRI-F fragment of adenovirus 2 genome. Gene. 1979 May;6(1):1–22. doi: 10.1016/0378-1119(79)90081-7. [DOI] [PubMed] [Google Scholar]
  12. 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]
  13. 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]
  14. 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]
  15. 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]
  16. Kruijer W., van Schaik F. M., Sussenbach J. S. Nucleotide sequence analysis of a region of adenovirus 5 DNA encoding a hitherto unidentified gene. Nucleic Acids Res. 1980 Dec 20;8(24):6033–6042. doi: 10.1093/nar/8.24.6033. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Levine A. J., Van Der Vliet P. C., Rosenwirth B., Rabek J., Frenkel G., Ensinger M. Adenovirus-infected, cell-specific, DNA-binding proteins. Cold Spring Harb Symp Quant Biol. 1975;39(Pt 1):559–566. doi: 10.1101/sqb.1974.039.01.069. [DOI] [PubMed] [Google Scholar]
  18. Levinson A. D., Postel E. H., Levine A. J. In vivo and in vitro phosphorylation of the adenovirus type 5 single strand-specific DNA-binding protein. Virology. 1977 Jun 1;79(1):144–159. doi: 10.1016/0042-6822(77)90341-5. [DOI] [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. Linné T., Jörnvall H., Philipson L. Purification and characterization of the phosphorylated DNA-binding protein from adenovirus-type-2-infected cells. Eur J Biochem. 1977 Jun 15;76(2):481–490. doi: 10.1111/j.1432-1033.1977.tb11618.x. [DOI] [PubMed] [Google Scholar]
  21. Linné T., Philipson L. Further characterization of the phosphate moiety of the adenovirus type 2 DNA-binding protein. Eur J Biochem. 1980 Jan;103(2):259–270. doi: 10.1111/j.1432-1033.1980.tb04310.x. [DOI] [PubMed] [Google Scholar]
  22. 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]
  23. Mayer A. J., Ginsberg H. S. Persistence of type 5 adenovirus DNA in cells transformed by temperature-sensitive mutant, H5ts125. Proc Natl Acad Sci U S A. 1977 Feb;74(2):785–788. doi: 10.1073/pnas.74.2.785. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. 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]
  25. Proudfoot N. J., Brownlee G. G. 3' non-coding region sequences in eukaryotic messenger RNA. Nature. 1976 Sep 16;263(5574):211–214. doi: 10.1038/263211a0. [DOI] [PubMed] [Google Scholar]
  26. Schechter N. M., Davies W., Anderson C. W. Adenovirus coded deoxyribonucleic acid binding protein. Isolation, physical properties, and effects of proteolytic digestion. Biochemistry. 1980 Jun 10;19(12):2802–2810. doi: 10.1021/bi00553a041. [DOI] [PubMed] [Google Scholar]
  27. Seif I., Khoury G., Dhar R. BKV splice sequences based on analysis of preferred donor and acceptor sites. Nucleic Acids Res. 1979 Jul 25;6(10):3387–3398. doi: 10.1093/nar/6.10.3387. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Sharp P. A. Speculations on RNA splicing. Cell. 1981 Mar;23(3):643–646. doi: 10.1016/0092-8674(81)90425-6. [DOI] [PubMed] [Google Scholar]
  29. Smith D. R., Calvo J. M. Nucleotide sequence of the E coli gene coding for dihydrofolate reductase. Nucleic Acids Res. 1980 May 24;8(10):2255–2274. doi: 10.1093/nar/8.10.2255. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Tibbetts C., Pettersson U., Johansson K., Philpson L. Relationship of mRNA from productively infected cells to the complementary strands of adenovirus type 2 DNA. J Virol. 1974 Feb;13(2):370–377. doi: 10.1128/jvi.13.2.370-377.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. 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]
  32. Van der Vliet P. C., Zandberg J., Jansz H. S. Evidence for a function of the adenovirus DNA-binding protein in initiation in DNA synthesis as well as in elongation of nascent DNA chains. Virology. 1977 Jul 1;80(1):98–110. doi: 10.1016/0042-6822(77)90383-x. [DOI] [PubMed] [Google Scholar]
  33. Vliet P. C., Sussenbach J. S. An adenovirus type 5 gene function required for initiation of viral DNA replication. Virology. 1975 Oct;67(2):415–426. doi: 10.1016/0042-6822(75)90443-2. [DOI] [PubMed] [Google Scholar]
  34. van der Vliet P. C., Keegstra W., Jansz H. S. Complex formation between the adenovirus type 5 DNA-binding protein and single-stranded DNA. Eur J Biochem. 1978 May 16;86(2):389–398. doi: 10.1111/j.1432-1033.1978.tb12321.x. [DOI] [PubMed] [Google Scholar]
  35. 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]

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