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. 1986 May;58(2):271–280. doi: 10.1128/jvi.58.2.271-280.1986

Parvovirus H-1 expression: mapping of the abundant cytoplasmic transcripts and identification of promoter sites and overlapping transcription units.

R M Lebovitz, R G Roeder
PMCID: PMC252910  PMID: 3009844

Abstract

The 5.2-kilobase (kb) genome of the autonomous parvovirus H-1 was transcribed in the rightward direction, yielding steady-state polyadenylated transcripts of 4.8, 3.2, and 2.9 kb. Detailed mapping of these transcripts demonstrated that the H-1 genome contained two overlapping transcription units: the larger unit extended from 4 map units (5' end) to 96 map units (3' end), and the smaller unit extended from 40 map units (5' end) to 96 map units (3' end). The 4.8- and 3.2-kb transcripts were derived from the larger transcription unit and differed by a 1,500-nucleotide segment (10 to 40 map units) which was present in the 4.8-kb transcript but was spliced from the 3.2-kb transcript. The 2.9-kb transcript, the most abundant of the three known H-1 transcripts, was derived from the smaller transcription unit. The sequence at each initiation site was consistent with the presence of a class II (RNA polymerase II) promoter, and cell-free transcription of parvovirus H-1 restriction fragments containing either promoter resulted in transcription of the correct DNA strand and produced 5' ends identical to those seen in vivo. All three transcripts contained a small but heterogeneous splice at 45 to 47 map units. Minor differences in splicing at this site may result in the synthesis of different viral proteins.

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Selected References

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

  1. Astell C. R., Thomson M., Merchlinsky M., Ward D. C. The complete DNA sequence of minute virus of mice, an autonomous parvovirus. Nucleic Acids Res. 1983 Feb 25;11(4):999–1018. doi: 10.1093/nar/11.4.999. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Berk A. J., Sharp P. A. Sizing and mapping of early adenovirus mRNAs by gel electrophoresis of S1 endonuclease-digested hybrids. Cell. 1977 Nov;12(3):721–732. doi: 10.1016/0092-8674(77)90272-0. [DOI] [PubMed] [Google Scholar]
  3. Bloom M. E., Race R. E., Wolfinbarger J. B. Identification of a nonvirion protein of Aleutian disease virus: mink with Aleutian disease have antibody to both virion and nonvirion proteins. J Virol. 1982 Aug;43(2):608–616. doi: 10.1128/jvi.43.2.608-616.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Breathnach R., Benoist C., O'Hare K., Gannon F., Chambon P. Ovalbumin gene: evidence for a leader sequence in mRNA and DNA sequences at the exon-intron boundaries. Proc Natl Acad Sci U S A. 1978 Oct;75(10):4853–4857. doi: 10.1073/pnas.75.10.4853. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Breathnach R., Chambon P. Organization and expression of eucaryotic split genes coding for proteins. Annu Rev Biochem. 1981;50:349–383. doi: 10.1146/annurev.bi.50.070181.002025. [DOI] [PubMed] [Google Scholar]
  6. Clinton G. M., Hayashi M. The parovivirus MVM: particles with altered structural proteins. Virology. 1975 Jul;66(1):261–261. doi: 10.1016/0042-6822(75)90196-8. [DOI] [PubMed] [Google Scholar]
  7. Cotmore S. F., Sturzenbecker L. J., Tattersall P. The autonomous parvovirus MVM encodes two nonstructural proteins in addition to its capsid polypeptides. Virology. 1983 Sep;129(2):333–343. doi: 10.1016/0042-6822(83)90172-1. [DOI] [PubMed] [Google Scholar]
  8. Efstratiadis A., Posakony J. W., Maniatis T., Lawn R. M., O'Connell C., Spritz R. A., DeRiel J. K., Forget B. G., Weissman S. M., Slightom J. L. The structure and evolution of the human beta-globin gene family. Cell. 1980 Oct;21(3):653–668. doi: 10.1016/0092-8674(80)90429-8. [DOI] [PubMed] [Google Scholar]
  9. Glisin V., Crkvenjakov R., Byus C. Ribonucleic acid isolated by cesium chloride centrifugation. Biochemistry. 1974 Jun 4;13(12):2633–2637. doi: 10.1021/bi00709a025. [DOI] [PubMed] [Google Scholar]
  10. Green M. R., Lebovitz R. M., Roeder R. G. Expression of the autonomous parvovirus H1 genome: evidence for a single transcriptional unit and multiple spliced polyadenylated transcripts. Cell. 1979 Aug;17(4):967–977. doi: 10.1016/0092-8674(79)90336-2. [DOI] [PubMed] [Google Scholar]
  11. Green M. R., Roeder R. G. Definition of a novel promoter for the major adenovirus-associated virus mRNA. Cell. 1980 Nov;22(1 Pt 1):231–242. doi: 10.1016/0092-8674(80)90171-3. [DOI] [PubMed] [Google Scholar]
  12. Green M. R., Roeder R. G. Transcripts of the adeno-associated virus genome: mapping of the major RNAs. J Virol. 1980 Oct;36(1):79–92. doi: 10.1128/jvi.36.1.79-92.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Hentschel C., Irminger J. C., Bucher P., Birnstiel M. L. Sea urchin histone mRNA termini are located in gene regions downstream from putative regulatory sequences. Nature. 1980 May 15;285(5761):147–151. doi: 10.1038/285147a0. [DOI] [PubMed] [Google Scholar]
  14. Hirt B. Selective extraction of polyoma DNA from infected mouse cell cultures. J Mol Biol. 1967 Jun 14;26(2):365–369. doi: 10.1016/0022-2836(67)90307-5. [DOI] [PubMed] [Google Scholar]
  15. Johnson F. B., Thomson T. A., Taylor P. A., Vlazny D. A. Molecular similarities among the adenovirus-associated virus polypeptides and evidence for a precursor protein. Virology. 1977 Oct 1;82(1):1–13. doi: 10.1016/0042-6822(77)90027-7. [DOI] [PubMed] [Google Scholar]
  16. Kongsvik J. R., Gierthy J. F., Rhode S. L., 3rd Replication process of the parvovirus H-1. IV. H-1-specific proteins synthesized in synchronized human NB kidney cells. J Virol. 1974 Dec;14(6):1600–1603. doi: 10.1128/jvi.14.6.1600-1603.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Lusby E. W., Berns K. I. Mapping of the 5' termini of two adeno-associated virus 2 RNAs in the left half of the genome. J Virol. 1982 Feb;41(2):518–526. doi: 10.1128/jvi.41.2.518-526.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Marcus C. J., Laughlin C. A., Carter B. J. Adeno-associated virus RNA transcription in vivo. Eur J Biochem. 1981 Dec;121(1):147–154. doi: 10.1111/j.1432-1033.1981.tb06443.x. [DOI] [PubMed] [Google Scholar]
  19. McMaster G. K., Carmichael G. G. Analysis of single- and double-stranded nucleic acids on polyacrylamide and agarose gels by using glyoxal and acridine orange. Proc Natl Acad Sci U S A. 1977 Nov;74(11):4835–4838. doi: 10.1073/pnas.74.11.4835. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Paradiso P. R., Williams K. R., Costantino R. L. Mapping of the amino terminus of the H-1 parvovirus major capsid protein. J Virol. 1984 Oct;52(1):77–81. doi: 10.1128/jvi.52.1.77-81.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Pintel D., Dadachanji D., Astell C. R., Ward D. C. The genome of minute virus of mice, an autonomous parvovirus, encodes two overlapping transcription units. Nucleic Acids Res. 1983 Feb 25;11(4):1019–1038. doi: 10.1093/nar/11.4.1019. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Pintel D., Merchlinsky M. J., Ward D. C. Expression of minute virus of mice structural proteins in murine cell lines transformed by bovine papillomavirus-minute virus of mice plasmid chimera. J Virol. 1984 Nov;52(2):320–327. doi: 10.1128/jvi.52.2.320-327.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Rhode S. L., 3rd, Paradiso P. R. Parvovirus genome: nucleotide sequence of H-1 and mapping of its genes by hybrid-arrested translation. J Virol. 1983 Jan;45(1):173–184. doi: 10.1128/jvi.45.1.173-184.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Rhode S. L., 3rd Replication process of the parvovirus H-1. II. Isolation and characterization of H-1 replicative form DNA. J Virol. 1974 Feb;13(2):400–410. doi: 10.1128/jvi.13.2.400-410.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Shenk T. E., Rhodes C., Rigby P. W., Berg P. Biochemical method for mapping mutational alterations in DNA with S1 nuclease: the location of deletions and temperature-sensitive mutations in simian virus 40. Proc Natl Acad Sci U S A. 1975 Mar;72(3):989–993. doi: 10.1073/pnas.72.3.989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Tattersall P., Shatkin A. J., Ward D. C. Sequence homology between the structural polypeptides of minute virus of mice. J Mol Biol. 1977 Apr 25;111(4):375–394. doi: 10.1016/s0022-2836(77)80060-0. [DOI] [PubMed] [Google Scholar]
  27. Vogelstein B., Gillespie D. Preparative and analytical purification of DNA from agarose. Proc Natl Acad Sci U S A. 1979 Feb;76(2):615–619. doi: 10.1073/pnas.76.2.615. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Weaver R. F., Weissmann C. Mapping of RNA by a modification of the Berk-Sharp procedure: the 5' termini of 15 S beta-globin mRNA precursor and mature 10 s beta-globin mRNA have identical map coordinates. Nucleic Acids Res. 1979 Nov 10;7(5):1175–1193. doi: 10.1093/nar/7.5.1175. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Weil P. A., Luse D. S., Segall J., Roeder R. G. Selective and accurate initiation of transcription at the Ad2 major late promotor in a soluble system dependent on purified RNA polymerase II and DNA. Cell. 1979 Oct;18(2):469–484. doi: 10.1016/0092-8674(79)90065-5. [DOI] [PubMed] [Google Scholar]
  30. Ziff E. B., Evans R. M. Coincidence of the promoter and capped 5' terminus of RNA from the adenovirus 2 major late transcription unit. Cell. 1978 Dec;15(4):1463–1475. doi: 10.1016/0092-8674(78)90070-3. [DOI] [PubMed] [Google Scholar]
  31. Zimmerman S. B., Sandeen D. The ribonuclease activity of crystallized pancreatic deoxyribonuclease. Anal Biochem. 1966 Feb;14(2):269–277. doi: 10.1016/0003-2697(66)90137-0. [DOI] [PubMed] [Google Scholar]

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