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An infectious cDNA copy of the genome of a non-cardiovirulent coxsackievirus B3 strain: its complete sequence analysis and comparison to the genomes of cardiovirulent coxsackieviruses

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Summary

The genome of the non-cardiovirulent coxsackievirus B3 (CVB3) strain CVB3/0 was cloned and sequenced to aid in the elucidation of the viral genetic basis for the CVB3 cardiovirulent phenotype. Reverse-transcribed sub-genomic complementary DNA (cDNA) fragments were enzymatically amplified using generic oligonucleotide primers and were assembled as a complete infectious genomic copy (pCVB3-0) downstream of the T7 RNA polymerase promoter. Positive-strand viral RNA transcribed from pCVB3-0 using T7 RNA polymerase and transfected into HeLa cells produced infectious virus (CVB3/0c). No differences in phenotype were observed comparing growth of CVB3/0c to the parental CVB3/0 in HeLa single-step growth curves, virus yields, or plaque size. When inoculated into C3H/HeJ mice. CVB3/0c achieved cardiac titers equivalent to the parental CVB3/0 and like the parental virus, demonstrated a non-cardiovirulent phenotype. The nucleotide sequence of the cloned CVB3/0 genome was determined and compared to the genomes of infectious cDNA clones of cardiovirulent CVB3 strains. Two consistent differences among nucleotides in non-translated regions and 8 amino acid differences relative to two well-characterized infectious cDNA copies of genomes from cardiovirulent CVB3 strains were identified.

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References

  1. Agol VI (1991) The 5'-untranslated region of picornaviral genomes. Adv Virus Res 40: 103–180

    Google Scholar 

  2. Almond JW (1987) The attenuation of poliovirus neurovirulence. Annu Rev Microbiol 41: 153–180

    Google Scholar 

  3. Beck MA, Tracy SM (1989) Murine cell-mediated immune response recognizes an enterovirus group-specific antigen(s). J Virol 63: 4148–4156

    Google Scholar 

  4. Bienz K, Egger D, Pfister T, Troxler M (1992) Structural and functional characterization of the poliovirus replication complex. J Virol 66: 2740–2747

    Google Scholar 

  5. Burch GJ, Sun S-H, Chu K-C, Sohal RS, Colcolough HL (1968) Interstitial and coxsackievirus B myocarditis in infants and children. JAMA 203: 55–62

    Google Scholar 

  6. Cherry JD (1987) Enteroviruses: polioviruses (poliomyelitis), coxsackieviruses, echoviruses, and enteroviruses. In: Feigin RD, Cherry JD (eds) Textbook of pediatric infectious diseases. Saunders, Philadelphia, pp. 1729–1790

    Google Scholar 

  7. del Angel RM, Papavassiliou AG, Fernandez-Tomas C, Silverstein SJ, Racaniello VR (1989) Cell proteins bind to multiple sites within the 5' untranslated region. Proc Natl Acad Sci USA 86: 8299–8303

    Google Scholar 

  8. Duke GM, Osorio JE, Palmenberg AC (1990) Attenuation of Mengo virus through genetic engineering of the 5' noncoding poly (C) tract. Nature 343: 474–476

    Google Scholar 

  9. Etchison D, Milburn SC, Edery I, Sonenberg N, Hershey JWB (1982) Inhibition of HeLa cell protein synthesis following poliovirus infection correlates with the proteolysis of a 220 000-dalton polypeptide associated with eucaryotic initiation factor 3 and a cap binding protein complex. J Biol Chem 257: 14806–14810

    Google Scholar 

  10. Gauntt CJ, Godeny EK, Lutton CW, Arizpe HM, Chapman NM, Tracy SM, Revtyak GE, Valente AJ, Rozek MM (1989) Mechanism(s) of coxsackievirus-induced acute myocarditis in the mouse In: de la Maza LM, Peterson EM (eds) Medical virology 8. Plenum, New York, pp 161–182

    Google Scholar 

  11. Gauntt CJ, Trousdale MD, LaBadie DRL, Paque RE, Nealon T (1979) Properties of coxsackievirus B3 variants which are amyocarditic or myocarditic for mice. J Med Virol 3: 207–220

    Google Scholar 

  12. Gauntt CJ, Trousdale MD, Lee JC, Paque RE (1983) Preliminary characterization of coxsackievirus B3 temperature-sensitive mutants. J Virol 45: 1037–1047

    Google Scholar 

  13. Filman DJ, Syed R, Chow M, Macadam AJ, Minor PD, Hogle JM (1989) Structural factors that control conformational transitions and serotype specificity in type 3 poliovirus. EMBO J 8: 1567–1579

    Google Scholar 

  14. Hogle JM, Chow M, Filman JD (1985) The three-dimensional structure of poliovirus at 2.9 A resolution. Science 229: 1358–1365

    Google Scholar 

  15. Innis M, Gelfand D (1990) Optimization of PCRs. In: Innis M, Gelfand D, Sninsky J, White T (eds) PCR protocols. Academic Press, San Diego, pp 3–12

    Google Scholar 

  16. Jacobson SJ, Konings DAM, Sarnow P (1993) Biochemcial and genetic evidence for a pseudoknot structure at the 3' terminus of the poliovirus RNA genome and its role in viral RNA amplification. J Virol 67: 2961–2971

    Google Scholar 

  17. Klump WM, Bergmann I, Muller BC, Ameis D, Kandolf R (1990) Complete nucleotide sequence of infectious coxsackievirus B3 cDNA: two initial 5' uridine residues are regained during plus-strand RNA synthesis. J Virol 64: 1573–1583

    Google Scholar 

  18. Krawczak M, Reiss J, Schmidtke J, Roesler U (1989) Polymerase chain reaction replication errors and reliability of gene diagnosis. Nucleic Acids Res 17: 2197–2201

    Google Scholar 

  19. Kuhn RJ, Wimmer E, Semler BL (1987) Expression of the poliovirus genome from infectious cDNA is dependent upon arrangements of eukaryotic and prokaryotic sequences in recombinant plasmids. Virology 157: 560–564

    Google Scholar 

  20. Leslie K, Blay R, Haisch V, Lodge A, Weller A, Huber S (1989) Clinical and experimental aspects of viral myocarditis. Clin Microbiol Rev 2: 191–203

    Google Scholar 

  21. Lindberg AM, Stalhandske POK, Pettersson U (1987) Genome of coxsackievirus B3. Virology 156: 50–63

    Google Scholar 

  22. Macadam AJ, Ferguson G, Arnold C, Minor PD (1991) An assembly defect as a result of an attenuating mutation in the capsid proteins of the poliovirus type 3 vaccine strain. J Virol 65: 5225–5231

    Google Scholar 

  23. Macadam AJ, Pollard SR, Ferguson G, Dunn G, Skuce R, Almond JW, Minor PD (1990) The 5' non-coding region of the type 2 poliovirus vaccine strain contains determinants of attenuation and temperature-sensitivity. Virology 181: 451–458

    Google Scholar 

  24. Martin A, Wychowski C, Couderic T, Crainic R, Hogle J, Girard M (1988) Engineering a poliovirus type 2 antigenic site on a type 1 capsid results in a chimeric virus which is neurovirulent for mice. EMBO J 7: 2839–2847

    Google Scholar 

  25. Meerovitch K, Nicholson R, Sonenberg N (1991) In vitro mutational analysis ofcis-acting RNA translational elements within the poliovirus type 2 5' untranslated region. J Virol 65: 5895–5901

    Google Scholar 

  26. Minor PD, Dunn G, Evans DMA, Magrath DI, John A, Howlett J, Phillips A, Westrop G, Wareham K, Almond JW, Hogle JM (1989) The temperature-sensitivity of the Sabin type 3 vaccine strain of poliovirus: molecular and structural effects of a mutation in the capsid protein VP3. J Gen Virol 70: 1117–1123

    Google Scholar 

  27. Minor PD, Ferguson M, Evans DMA, Almond JW, Icenogle JP (1986) Antigenic structure of polioviruses of serotypes 1, 2, 3. J Gen Virol 67: 1283–1291

    Google Scholar 

  28. Moscufo N, Chow M (1992) Myristate-protein interactions in poliovirus: interactions of VP4 threonine 28 contribute to the structural conformation of assembly intermediates and the stability of assembled virions. J Virol 66: 6849–6857

    Google Scholar 

  29. Olsen EG (1992) The pathogenesis of dilated cardiomyopathy. Postgrad Med J 68 [Suppl 1]: S7–10

    Google Scholar 

  30. Palmenberg AC (1989) Sequence alignments of picornaviral capsids. In: Semler BL, Ehrenfeld E (eds) Molecular aspects of picornavirus infection and detection. American Society for Microbiology, Washington, pp 211–242

    Google Scholar 

  31. Pestova TV, Hellen CUT, Wimmer E (1991) Translation of poliovirus RNA: role of an essentialcis-acting oligopyrimidine element within the 5' nontranslated region and involvement of a cellular 57-kilodalton protein. J Virol 65: 6194–6204

    Google Scholar 

  32. Pilipenko EV, Maslova SV, Sinyakov AN, Agol VI (1992) Towards identification ofcis-acting elements involved in the replication of enterovirus and rhinovirus RNAs: a proposal for the existence of tRNA-like terminal structures. Nucleic Acids Res 20: 1739–1745

    Google Scholar 

  33. Ramsingh A, Hixson A, Duceman B, Slack J (1990) Evidence suggesting that virulence maps to the P1 region of the coxsackie-virus B4 genome. J Virol 64: 3078–3081

    Google Scholar 

  34. Ramsingh A, Araki H, Bryant S, Hixson A (1992) Identification of candidate sequences that determine virulence in coxsackievirus B4. Virus Res 23: 281–292

    Google Scholar 

  35. Reimann BY, Zell R, Kandolf R (1991) Mapping of a neutralizing antigenic site of coxsackievirus B4 by construction of an antigen chimera. J Virol 65: 3475–3480

    Google Scholar 

  36. Ren R, Moss EG, Racaniello VR (1991) Identification of two determinants that attenuate vaccine-related type 2 poliovirus. J Virol 65: 1377–1382

    Google Scholar 

  37. Rodriguez M, Roos RP (1992) Pathogenesis of early and late disease in mice infected with Theiler's virus, using intratypic recombinant GDVII/DA viruses. J Virol 66: 217–225

    Google Scholar 

  38. Rose NR, Neumann DA, Herskowitz A, Traystman MD (1988) Genetics of susceptibility to viral myocarditis in mice. Pathol Immunopathol Res 7: 266–278

    Google Scholar 

  39. Rossman MG, Arnold E, Erickson JW, Frankenberger EA, Griffith AG, Hecht HJ, Johnson JE, Kamer G, Luo M, Mosser AG, Rueckert RR, Sherry B, Vriend G (1985) Structure of a human common cold virus and functional relationship to other picornaviruses. Nature 317: 145–153

    Google Scholar 

  40. Rueckert R, Wimmer E (1984) Systematic nomenclature of picornavirus proteins. J Virol 50: 957–959

    Google Scholar 

  41. Sarnow P, Bernstein HD, Baltimore D (1986) A poliovirus temperature-sensitive RNA synthesis mutant located in a noncoding region of the genome. Proc Natl Acad Sci USA 83: 571–575

    Google Scholar 

  42. Skinner MA, Racaniello VR, Dunn G, Cooper J, Minor PD, Almond JW (1989) New model for the secondary structure of the 5' non-coding RNA of poliovirus is supported by biochemical and genetic data that also show that RNA secondary structure is important in neurovirulence. J Mol Biol 207: 379–392

    Google Scholar 

  43. Stanway G (1990) Structure, function and evolution of picornaviruses. J Gen Virol 71: 2483–2501

    Google Scholar 

  44. Tabor S, Richardson CC (1987) DNA sequence analysis with a modified bacteriophage T7 DNA polymerase. Proc Natl Acad Sci USA 84: 4767–4771

    Google Scholar 

  45. Tangy F, McAllister A, Aubert C, Brahic M (1991) Determinants of persistence and demyelination of the DA Strain of Theiler's virus are found only in the VP1 gene. J Virol 65: 1616–1618

    Google Scholar 

  46. Tindall KR, Kunkel TA (1988) Fidelity of DNA synthesis by the Thermus aquaticus DNA polymerase. Biochemistry 27: 6008–6013

    Google Scholar 

  47. Tracy S (1985) Comparison of genomic homologies in the coxsackievirus B group by use of cDNA: RNA dot-blot hybridization. J Clin Microbiol 21: 371–374

    Google Scholar 

  48. Tracy S, Chapman NM, Beck MA (1991) Molecular biology and pathogenesis of coxsackie B viruses. Rev Med Virol 1: 145–154

    Google Scholar 

  49. Tracy S, Chapman N, Pallansch M, Coller B, Beck M, Kolbeck M, Woronoff Y (1991) Characterization and molecular cloning of a coxsackie B virus from a human myocarditic heart. J Cell Biochem 15E: M342

  50. Tracy S, Chapman NM, Pistillo JM (1992) Detection of human enteroviruses using the polymerase chain reaction. In: Becker Y, Darai G (eds) Frontiers of virology. Springer, Berlin, Heidelberg, New York, Tokyo, pp 331–344

    Google Scholar 

  51. Tracy S, Chapman NM, Tu Z (1992) Coxsackievirus B3 from an infectious cDNA copy of the genome is cardiovirulent in mice. Arch Virol 122: 398–409

    Google Scholar 

  52. Trousdale MD, Paque RE, Gauntt CJ (1977) Isolation of coxsackievirus B3 temperature-sensitive mutants and their assignment to complementation groups. Biochem Biophys Res Commun 76: 368–375

    Google Scholar 

  53. van der Werf S, Bradley J, Wimmer E, Studier FW, Dunn JJ (1986) Synthesis of infectious poliovirus RNA by purified T7 RNA polymerase. Proc Natl Acad Sci USA 83: 2330–2334

    Google Scholar 

  54. Woodruff JF (1980) Viral myocarditis: a review. Am J Pathol 101: 425–483

    Google Scholar 

  55. Woodruff JF, Kilbourne ED (1970) The influence of quantitative post weaning undernutrition of coxsackievirus B3 infection of adult mice. I. Viral persistence and increased severity of lesions. J Infect Dis 121: 137–163

    Google Scholar 

  56. Yeates TO, Jacobson DH, Martin A, Wychowski C, Girard M, Filman DJ, Hogle JM (1991) Three-dimensional structure of a mouse-adapted type 2/type 1 poliovirus chimera. EMBO J 10: 2331–2341

    Google Scholar 

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Authors and Affiliations

  1. Department of Pathology and Microbiology, University of Nebraska Medical Center, Omaha, Nebraska

    Nora M. Chapman, Z. Tu & S. Tracy

  2. Department of Microbiology, University of Texas Health Science Center, San Antonio, Texas, USA

    C. J. Gauntt

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  1. Nora M. Chapman
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  2. Z. Tu
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  4. C. J. Gauntt
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Chapman, N.M., Tu, Z., Tracy, S. et al. An infectious cDNA copy of the genome of a non-cardiovirulent coxsackievirus B3 strain: its complete sequence analysis and comparison to the genomes of cardiovirulent coxsackieviruses. Archives of Virology 135, 115–130 (1994). https://doi.org/10.1007/BF01309769

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  • DOI: https://doi.org/10.1007/BF01309769

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