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. 1993 Aug;67(8):4630–4638. doi: 10.1128/jvi.67.8.4630-4638.1993

A mutation in the RNA polymerase of poliovirus type 1 contributes to attenuation in mice.

M Tardy-Panit 1, B Blondel 1, A Martin 1, F Tekaia 1, F Horaud 1, F Delpeyroux 1
PMCID: PMC237848  PMID: 8392604

Abstract

The attenuated Sabin strain of poliovirus type 1 (PV-1) differs from the neurovirulent PV-1 Mahoney strain by 55 nucleotide mutations. Only one of these mutations (A-480-->G, in the 5' noncoding (5' NC) region of the genome, is well characterized, and it confers a strong attenuating effect. We attempted to identify genetic attenuation determinants in the 3'-terminal part of the Sabin 1 genome including the 3D polymerase (3Dpol) gene and the 3' NC region. Previous studies suggested that some of the 11 mutations in this region of the Sabin 1 genome, and in particular a mutation in the polymerase gene (U-6203-->C, Tyr-73-->His), are involved to some extent in the attenuation of PV-1. We analyzed the attenuating effect in the mouse model by using the mouse-adapted PV-1/PV-2 chimeric strain v510 (a Mahoney strain carrying nine amino acids of the VP1 capsid protein from the Lansing strain of PV-2). Mutagenesis of locus 6203 was performed on the original v510 (U-6203-->C) and also on a hybrid v510/Sabin 1 (C-6203-->U) carrying the downstream 1,840 nucleotides of the Sabin 1 genome including the 3Dpol and 3' NC regions. Statistical analysis of disease incidence and time to disease onset in numerous mice inoculated with these strains strongly suggested that nucleotide C-6203 is involved in the attenuation of the Sabin 1 strain. Results also suggested that, among the mutations located in the 3Dpol and 3' NC regions, nucleotide C-6203 may be the principal or the only one to be involved in attenuation in this mouse model. We also found that the effect of C-6203 was weaker than that of nucleotide G-480; the two nucleotides acted independently and may have a cumulative effect on attenuation. The U-6203-->C substitution also appeared to contribute to the thermosensitivity of the Sabin 1 strain.

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Selected References

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  1. Agol V. I., Drozdov S. G., Grachev V. P., Kolesnikova M. S., Kozlov V. G., Ralph N. M., Romanova L. I., Tolskaya E. A., Tyufanov A. V., Viktorova E. G. Recombinants between attenuated and virulent strains of poliovirus type 1: derivation and characterization of recombinants with centrally located crossover points. Virology. 1985 Jun;143(2):467–477. doi: 10.1016/0042-6822(85)90386-1. [DOI] [PubMed] [Google Scholar]
  2. Agut H., Kean K. M., Fichot O., Morasco J., Flanegan J. B., Girard M. A point mutation in the poliovirus polymerase gene determines a complementable temperature-sensitive defect of RNA replication. Virology. 1989 Feb;168(2):302–311. doi: 10.1016/0042-6822(89)90270-5. [DOI] [PubMed] [Google Scholar]
  3. Andino R., Rieckhof G. E., Baltimore D. A functional ribonucleoprotein complex forms around the 5' end of poliovirus RNA. Cell. 1990 Oct 19;63(2):369–380. doi: 10.1016/0092-8674(90)90170-j. [DOI] [PubMed] [Google Scholar]
  4. BODIAN D. Emerging concept of poliomyelitis infection. Science. 1955 Jul 15;122(3159):105–108. doi: 10.1126/science.122.3159.105. [DOI] [PubMed] [Google Scholar]
  5. Balanant J., Guillot S., Candrea A., Delpeyroux F., Crainic R. The natural genomic variability of poliovirus analyzed by a restriction fragment length polymorphism assay. Virology. 1991 Oct;184(2):645–654. doi: 10.1016/0042-6822(91)90434-d. [DOI] [PubMed] [Google Scholar]
  6. Bernstein H. D., Sarnow P., Baltimore D. Genetic complementation among poliovirus mutants derived from an infectious cDNA clone. J Virol. 1986 Dec;60(3):1040–1049. doi: 10.1128/jvi.60.3.1040-1049.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Burns C. C., Lawson M. A., Semler B. L., Ehrenfeld E. Effects of mutations in poliovirus 3Dpol on RNA polymerase activity and on polyprotein cleavage. J Virol. 1989 Nov;63(11):4866–4874. doi: 10.1128/jvi.63.11.4866-4874.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Burns C. C., Richards O. C., Ehrenfeld E. Temperature-sensitive polioviruses containing mutations in RNA polymerase. Virology. 1992 Aug;189(2):568–582. doi: 10.1016/0042-6822(92)90580-i. [DOI] [PubMed] [Google Scholar]
  9. Cammack N., Phillips A., Dunn G., Patel V., Minor P. D. Intertypic genomic rearrangements of poliovirus strains in vaccinees. Virology. 1988 Dec;167(2):507–514. [PubMed] [Google Scholar]
  10. Charini W. A., Burns C. C., Ehrenfeld E., Semler B. L. trans rescue of a mutant poliovirus RNA polymerase function. J Virol. 1991 May;65(5):2655–2665. doi: 10.1128/jvi.65.5.2655-2665.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Christodoulou C., Colbere-Garapin F., Macadam A., Taffs L. F., Marsden S., Minor P., Horaud F. Mapping of mutations associated with neurovirulence in monkeys infected with Sabin 1 poliovirus revertants selected at high temperature. J Virol. 1990 Oct;64(10):4922–4929. doi: 10.1128/jvi.64.10.4922-4929.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Colbère-Garapin F., Christodoulou C., Crainic R., Garapin A. C., Candrea A. Addition of a foreign oligopeptide to the major capsid protein of poliovirus. Proc Natl Acad Sci U S A. 1988 Nov;85(22):8668–8672. doi: 10.1073/pnas.85.22.8668. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Contreras G., Dimock K., Furesz J., Gardell C., Hazlett D., Karpinski K., McCorkle G., Wu L. Genetic characterization of Sabin types 1 and 3 poliovaccine virus following serial passage in the human intestinal tract. Biologicals. 1992 Mar;20(1):15–26. doi: 10.1016/s1045-1056(05)80003-x. [DOI] [PubMed] [Google Scholar]
  14. Couderc T., Christodoulou C., Kopecka H., Marsden S., Taffs L. F., Crainic R., Horaud F. Molecular pathogenesis of neural lesions induced by poliovirus type 1. J Gen Virol. 1989 Nov;70(Pt 11):2907–2918. doi: 10.1099/0022-1317-70-11-2907. [DOI] [PubMed] [Google Scholar]
  15. Dedieu J. F., Ronco J., van der Werf S., Hogle J. M., Henin Y., Girard M. Poliovirus chimeras expressing sequences from the principal neutralization domain of human immunodeficiency virus type 1. J Virol. 1992 May;66(5):3161–3167. doi: 10.1128/jvi.66.5.3161-3167.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Fichot O., Girard M. An improved method for sequencing of RNA templates. Nucleic Acids Res. 1990 Oct 25;18(20):6162–6162. doi: 10.1093/nar/18.20.6162. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Jablonski S. A., Luo M., Morrow C. D. Enzymatic activity of poliovirus RNA polymerase mutants with single amino acid changes in the conserved YGDD amino acid motif. J Virol. 1991 Sep;65(9):4565–4572. doi: 10.1128/jvi.65.9.4565-4572.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Jubelt B., Ropka S. L., Goldfarb S., Waltenbaugh C., Oates R. P. Susceptibility and resistance to poliovirus-induced paralysis of inbred mouse strains. J Virol. 1991 Feb;65(2):1035–1040. doi: 10.1128/jvi.65.2.1035-1040.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Kawamura N., Kohara M., Abe S., Komatsu T., Tago K., Arita M., Nomoto A. Determinants in the 5' noncoding region of poliovirus Sabin 1 RNA that influence the attenuation phenotype. J Virol. 1989 Mar;63(3):1302–1309. doi: 10.1128/jvi.63.3.1302-1309.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Kean K. M., Agut H., Fichot O., Girard M. Substitution in the poliovirus replicase gene determines actinomycin D sensitivity of viral replication at elevated temperature. Virus Res. 1989 Jan;12(1):19–32. doi: 10.1016/0168-1702(89)90050-6. [DOI] [PubMed] [Google Scholar]
  21. Kohara M., Abe S., Kuge S., Semler B. L., Komatsu T., Arita M., Itoh H., Nomoto A. An infectious cDNA clone of the poliovirus Sabin strain could be used as a stable repository and inoculum for the oral polio live vaccine. Virology. 1986 May;151(1):21–30. doi: 10.1016/0042-6822(86)90100-5. [DOI] [PubMed] [Google Scholar]
  22. Koike S., Taya C., Kurata T., Abe S., Ise I., Yonekawa H., Nomoto A. Transgenic mice susceptible to poliovirus. Proc Natl Acad Sci U S A. 1991 Feb 1;88(3):951–955. doi: 10.1073/pnas.88.3.951. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Kuge S., Nomoto A. Construction of viable deletion and insertion mutants of the Sabin strain of type 1 poliovirus: function of the 5' noncoding sequence in viral replication. J Virol. 1987 May;61(5):1478–1487. doi: 10.1128/jvi.61.5.1478-1487.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. La Monica N., Almond J. W., Racaniello V. R. A mouse model for poliovirus neurovirulence identifies mutations that attenuate the virus for humans. J Virol. 1987 Sep;61(9):2917–2920. doi: 10.1128/jvi.61.9.2917-2920.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. La Monica N., Racaniello V. R. Differences in replication of attenuated and neurovirulent polioviruses in human neuroblastoma cell line SH-SY5Y. J Virol. 1989 May;63(5):2357–2360. doi: 10.1128/jvi.63.5.2357-2360.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Lipskaya G. Y., Muzychenko A. R., Kutitova O. K., Maslova S. V., Equestre M., Drozdov S. G., Bercoff R. P., Agol V. I. Frequent isolation of intertypic poliovirus recombinants with serotype 2 specificity from vaccine-associated polio cases. J Med Virol. 1991 Dec;35(4):290–296. doi: 10.1002/jmv.1890350415. [DOI] [PubMed] [Google Scholar]
  27. Macadam A. J., Pollard S. R., Ferguson G., Dunn G., Skuce R., Almond J. W., Minor P. D. The 5' noncoding region of the type 2 poliovirus vaccine strain contains determinants of attenuation and temperature sensitivity. Virology. 1991 Apr;181(2):451–458. doi: 10.1016/0042-6822(91)90877-e. [DOI] [PubMed] [Google Scholar]
  28. Martin A., Benichou D., Couderc T., Hogle J. M., Wychowski C., Van der Werf S., Girard M. Use of type 1/type 2 chimeric polioviruses to study determinants of poliovirus type 1 neurovirulence in a mouse model. Virology. 1991 Feb;180(2):648–658. doi: 10.1016/0042-6822(91)90078-p. [DOI] [PubMed] [Google Scholar]
  29. Martin A., Wychowski C., Couderc T., Crainic R., Hogle J., Girard M. Engineering a poliovirus type 2 antigenic site on a type 1 capsid results in a chimaeric virus which is neurovirulent for mice. EMBO J. 1988 Sep;7(9):2839–2847. doi: 10.1002/j.1460-2075.1988.tb03140.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Moss E. G., O'Neill R. E., Racaniello V. R. Mapping of attenuating sequences of an avirulent poliovirus type 2 strain. J Virol. 1989 May;63(5):1884–1890. doi: 10.1128/jvi.63.5.1884-1890.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Murray M. G., Bradley J., Yang X. F., Wimmer E., Moss E. G., Racaniello V. R. Poliovirus host range is determined by a short amino acid sequence in neutralization antigenic site I. Science. 1988 Jul 8;241(4862):213–215. doi: 10.1126/science.2838906. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Muzychenko A. R., Lipskaya GYu, Maslova S. V., Svitkin Y. V., Pilipenko E. V., Nottay B. K., Kew O. M., Agol V. I. Coupled mutations in the 5'-untranslated region of the Sabin poliovirus strains during in vivo passages: structural and functional implications. Virus Res. 1991 Oct;21(2):111–122. doi: 10.1016/0168-1702(91)90002-d. [DOI] [PubMed] [Google Scholar]
  33. Nakano J. H., Hatch M. H., Thieme M. L., Nottay B. Parameters for differentiating vaccine-derived and wild poliovirus strains. Prog Med Virol. 1978;24:178–206. [PubMed] [Google Scholar]
  34. Nomoto A., Omata T., Toyoda H., Kuge S., Horie H., Kataoka Y., Genba Y., Nakano Y., Imura N. Complete nucleotide sequence of the attenuated poliovirus Sabin 1 strain genome. Proc Natl Acad Sci U S A. 1982 Oct;79(19):5793–5797. doi: 10.1073/pnas.79.19.5793. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Omata T., Kohara M., Kuge S., Komatsu T., Abe S., Semler B. L., Kameda A., Itoh H., Arita M., Wimmer E. Genetic analysis of the attenuation phenotype of poliovirus type 1. J Virol. 1986 May;58(2):348–358. doi: 10.1128/jvi.58.2.348-358.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Otelea D., Guillot S., Furione M., Combiescu A. A., Balanant J., Candrea A., Crainic R. Genomic modifications in naturally occurring neurovirulent revertants of Sabin 1 polioviruses. Dev Biol Stand. 1993;78:33–38. [PubMed] [Google Scholar]
  37. Pelletier J., Sonenberg N. Internal initiation of translation of eukaryotic mRNA directed by a sequence derived from poliovirus RNA. Nature. 1988 Jul 28;334(6180):320–325. doi: 10.1038/334320a0. [DOI] [PubMed] [Google Scholar]
  38. Pilipenko E. V., Blinov V. M., Romanova L. I., Sinyakov A. N., Maslova S. V., Agol V. I. Conserved structural domains in the 5'-untranslated region of picornaviral genomes: an analysis of the segment controlling translation and neurovirulence. Virology. 1989 Feb;168(2):201–209. doi: 10.1016/0042-6822(89)90259-6. [DOI] [PubMed] [Google Scholar]
  39. Racaniello V. R. Poliovirus neurovirulence. Adv Virus Res. 1988;34:217–246. doi: 10.1016/s0065-3527(08)60519-9. [DOI] [PubMed] [Google Scholar]
  40. Ren R. B., Costantini F., Gorgacz E. J., Lee J. J., Racaniello V. R. Transgenic mice expressing a human poliovirus receptor: a new model for poliomyelitis. Cell. 1990 Oct 19;63(2):353–362. doi: 10.1016/0092-8674(90)90168-e. [DOI] [PubMed] [Google Scholar]
  41. Ren R. B., Moss E. G., Racaniello V. R. Identification of two determinants that attenuate vaccine-related type 2 poliovirus. J Virol. 1991 Mar;65(3):1377–1382. doi: 10.1128/jvi.65.3.1377-1382.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Richards O. C., Yu P., Neufeld K. L., Ehrenfeld E. Nucleotide binding by the poliovirus RNA polymerase. J Biol Chem. 1992 Aug 25;267(24):17141–17146. [PubMed] [Google Scholar]
  43. Sarnow P., Bernstein H. D., Baltimore D. A poliovirus temperature-sensitive RNA synthesis mutant located in a noncoding region of the genome. Proc Natl Acad Sci U S A. 1986 Feb;83(3):571–575. doi: 10.1073/pnas.83.3.571. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. Sarnow P., Jacobson S. J., Najita L. Poliovirus genetics. Curr Top Microbiol Immunol. 1990;161:155–188. doi: 10.1007/978-3-642-75602-3_6. [DOI] [PubMed] [Google Scholar]
  45. Skinner M. A., Racaniello V. R., Dunn G., Cooper J., Minor P. D., Almond J. W. 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. 1989 May 20;207(2):379–392. doi: 10.1016/0022-2836(89)90261-1. [DOI] [PubMed] [Google Scholar]
  46. Strebel P. M., Sutter R. W., Cochi S. L., Biellik R. J., Brink E. W., Kew O. M., Pallansch M. A., Orenstein W. A., Hinman A. R. Epidemiology of poliomyelitis in the United States one decade after the last reported case of indigenous wild virus-associated disease. Clin Infect Dis. 1992 Feb;14(2):568–579. doi: 10.1093/clinids/14.2.568. [DOI] [PubMed] [Google Scholar]
  47. Svitkin Y. V., Cammack N., Minor P. D., Almond J. W. Translation deficiency of the Sabin type 3 poliovirus genome: association with an attenuating mutation C472----U. Virology. 1990 Mar;175(1):103–109. doi: 10.1016/0042-6822(90)90190-3. [DOI] [PubMed] [Google Scholar]
  48. Tatem J. M., Weeks-Levy C., Georgiu A., DiMichele S. J., Gorgacz E. J., Racaniello V. R., Cano F. R., Mento S. J. A mutation present in the amino terminus of Sabin 3 poliovirus VP1 protein is attenuating. J Virol. 1992 May;66(5):3194–3197. doi: 10.1128/jvi.66.5.3194-3197.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  49. Toyoda H., Kohara M., Kataoka Y., Suganuma T., Omata T., Imura N., Nomoto A. Complete nucleotide sequences of all three poliovirus serotype genomes. Implication for genetic relationship, gene function and antigenic determinants. J Mol Biol. 1984 Apr 25;174(4):561–585. doi: 10.1016/0022-2836(84)90084-6. [DOI] [PubMed] [Google Scholar]
  50. Toyoda H., Yang C. F., Takeda N., Nomoto A., Wimmer E. Analysis of RNA synthesis of type 1 poliovirus by using an in vitro molecular genetic approach. J Virol. 1987 Sep;61(9):2816–2822. doi: 10.1128/jvi.61.9.2816-2822.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  51. Westrop G. D., Wareham K. A., Evans D. M., Dunn G., Minor P. D., Magrath D. I., Taffs F., Marsden S., Skinner M. A., Schild G. C. Genetic basis of attenuation of the Sabin type 3 oral poliovirus vaccine. J Virol. 1989 Mar;63(3):1338–1344. doi: 10.1128/jvi.63.3.1338-1344.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]

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