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. 1994 Sep;68(9):5460–5468. doi: 10.1128/jvi.68.9.5460-5468.1994

Oligo(A) sequences of human respiratory syncytial virus G protein gene: assessment of their genetic stability in frameshift mutants.

B García-Barreno 1, T Delgado 1, J A Melero 1
PMCID: PMC236946  PMID: 8057428

Abstract

We have described previously antibody-resistant mutants of the human respiratory syncytial virus Long strain that contained frameshift changes generated by deletions or insertions of a single adenosine in oligo(A) tracts (mRNA sense) of the G protein gene. Since these mutations introduced drastic structural and antigenic changes in the G protein C-terminal third, we decided to test the mutant stability by passaging the viruses in either the presence or the absence of selective antibody. Two such mutants (R63/1/2/3 and R63/2/4/8), with a single reading frame shift, reverted after a few passages in the absence of antibody to the wild-type genotype, by insertion of an A at the same homopolymeric tract as in the original deletion. In contrast, a double frameshift mutant (R63/2/4/1), generated by deletion of an A after nucleotide 623 and insertion of another A seven triplets later, was stably maintained after passage in either the absence or the presence of antibody. The stability of this mutant was manifested in its capacity to gradually displace the Long strain from mixed infections and by the fact that mutant R63/2/4/8 acquired the genotype of R63/2/4/1 after several passages in the presence of antibody. These results were indicative of genetic instability in the oligo(A) tract length of certain G protein mutants, which resulted in frameshift changes. The frequency of such errors among the viral RNA population obtained from a single infectious cycle was estimated to be lower than 1%. The relevance of these results for respiratory syncytial virus evolution is 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. Anderson L. J., Hierholzer J. C., Tsou C., Hendry R. M., Fernie B. F., Stone Y., McIntosh K. Antigenic characterization of respiratory syncytial virus strains with monoclonal antibodies. J Infect Dis. 1985 Apr;151(4):626–633. doi: 10.1093/infdis/151.4.626. [DOI] [PubMed] [Google Scholar]
  2. Batschelet E., Domingo E., Weissmann C. The proportion of revertant and mutant phage in a growing population, as a function of mutation and growth rate. Gene. 1976;1(1):27–32. doi: 10.1016/0378-1119(76)90004-4. [DOI] [PubMed] [Google Scholar]
  3. Cane P. A., Matthews D. A., Pringle C. R. Frequent polymerase errors observed in a restricted area of clones derived from the attachment (G) protein gene of respiratory syncytial virus. J Virol. 1993 Feb;67(2):1090–1093. doi: 10.1128/jvi.67.2.1090-1093.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Cane P. A., Matthews D. A., Pringle C. R. Identification of variable domains of the attachment (G) protein of subgroup A respiratory syncytial viruses. J Gen Virol. 1991 Sep;72(Pt 9):2091–2096. doi: 10.1099/0022-1317-72-9-2091. [DOI] [PubMed] [Google Scholar]
  5. Cattaneo R., Schmid A., Eschle D., Baczko K., ter Meulen V., Billeter M. A. Biased hypermutation and other genetic changes in defective measles viruses in human brain infections. Cell. 1988 Oct 21;55(2):255–265. doi: 10.1016/0092-8674(88)90048-7. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Chirgwin J. M., Przybyla A. E., MacDonald R. J., Rutter W. J. Isolation of biologically active ribonucleic acid from sources enriched in ribonuclease. Biochemistry. 1979 Nov 27;18(24):5294–5299. doi: 10.1021/bi00591a005. [DOI] [PubMed] [Google Scholar]
  7. Collins P. L., Mottet G. Oligomerization and post-translational processing of glycoprotein G of human respiratory syncytial virus: altered O-glycosylation in the presence of brefeldin A. J Gen Virol. 1992 Apr;73(Pt 4):849–863. doi: 10.1099/0022-1317-73-4-849. [DOI] [PubMed] [Google Scholar]
  8. Garcia-Barreno B., Jorcano J. L., Aukenbauer T., López-Galíndez C., Melero J. A. Participation of cytoskeletal intermediate filaments in the infectious cycle of human respiratory syncytial virus (RSV). Virus Res. 1988 Mar;9(4):307–321. doi: 10.1016/0168-1702(88)90090-1. [DOI] [PubMed] [Google Scholar]
  9. García-Barreno B., Palomo C., Peñas C., Delgado T., Perez-Breña P., Melero J. A. Marked differences in the antigenic structure of human respiratory syncytial virus F and G glycoproteins. J Virol. 1989 Feb;63(2):925–932. doi: 10.1128/jvi.63.2.925-932.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. García-Barreno B., Portela A., Delgado T., López J. A., Melero J. A. Frame shift mutations as a novel mechanism for the generation of neutralization resistant mutants of human respiratory syncytial virus. EMBO J. 1990 Dec;9(12):4181–4187. doi: 10.1002/j.1460-2075.1990.tb07642.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Gonzalez M. J., Saiz J. C., Laor O., Moore D. M. Antigenic stability of foot-and-mouth disease virus variants on serial passage in cell culture. J Virol. 1991 Jul;65(7):3949–3953. doi: 10.1128/jvi.65.7.3949-3953.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Hendricks D. A., McIntosh K., Patterson J. L. Further characterization of the soluble form of the G glycoprotein of respiratory syncytial virus. J Virol. 1988 Jul;62(7):2228–2233. doi: 10.1128/jvi.62.7.2228-2233.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Ionov Y., Peinado M. A., Malkhosyan S., Shibata D., Perucho M. Ubiquitous somatic mutations in simple repeated sequences reveal a new mechanism for colonic carcinogenesis. Nature. 1993 Jun 10;363(6429):558–561. doi: 10.1038/363558a0. [DOI] [PubMed] [Google Scholar]
  14. Jacques J. P., Kolakofsky D. Pseudo-templated transcription in prokaryotic and eukaryotic organisms. Genes Dev. 1991 May;5(5):707–713. doi: 10.1101/gad.5.5.707. [DOI] [PubMed] [Google Scholar]
  15. Johnson P. R., Spriggs M. K., Olmsted R. A., Collins P. L. The G glycoprotein of human respiratory syncytial viruses of subgroups A and B: extensive sequence divergence between antigenically related proteins. Proc Natl Acad Sci U S A. 1987 Aug;84(16):5625–5629. doi: 10.1073/pnas.84.16.5625. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Kunkel T. A. Frameshift mutagenesis by eucaryotic DNA polymerases in vitro. J Biol Chem. 1986 Oct 15;261(29):13581–13587. [PubMed] [Google Scholar]
  17. Lambert D. M. Role of oligosaccharides in the structure and function of respiratory syncytial virus glycoproteins. Virology. 1988 Jun;164(2):458–466. doi: 10.1016/0042-6822(88)90560-0. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Levine S., Klaiber-Franco R., Paradiso P. R. Demonstration that glycoprotein G is the attachment protein of respiratory syncytial virus. J Gen Virol. 1987 Sep;68(Pt 9):2521–2524. doi: 10.1099/0022-1317-68-9-2521. [DOI] [PubMed] [Google Scholar]
  19. Linton M. F., Pierotti V., Young S. G. Reading-frame restoration with an apolipoprotein B gene frameshift mutation. Proc Natl Acad Sci U S A. 1992 Dec 1;89(23):11431–11435. doi: 10.1073/pnas.89.23.11431. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. López J. A., Villanueva N., Melero J. A., Portela A. Nucleotide sequence of the fusion and phosphoprotein genes of human respiratory syncytial (RS) virus Long strain: evidence of subtype genetic heterogeneity. Virus Res. 1988 May;10(2-3):249–261. doi: 10.1016/0168-1702(88)90020-2. [DOI] [PubMed] [Google Scholar]
  21. Martínez M. A., Carrillo C., González-Candelas F., Moya A., Domingo E., Sobrino F. Fitness alteration of foot-and-mouth disease virus mutants: measurement of adaptability of viral quasispecies. J Virol. 1991 Jul;65(7):3954–3957. doi: 10.1128/jvi.65.7.3954-3957.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Mufson M. A., Orvell C., Rafnar B., Norrby E. Two distinct subtypes of human respiratory syncytial virus. J Gen Virol. 1985 Oct;66(Pt 10):2111–2124. doi: 10.1099/0022-1317-66-10-2111. [DOI] [PubMed] [Google Scholar]
  23. Palomo C., García-Barreno B., Peñas C., Melero J. A. The G protein of human respiratory syncytial virus: significance of carbohydrate side-chains and the C-terminal end to its antigenicity. J Gen Virol. 1991 Mar;72(Pt 3):669–675. doi: 10.1099/0022-1317-72-3-669. [DOI] [PubMed] [Google Scholar]
  24. Park K. H., Krystal M. In vivo model for pseudo-templated transcription in Sendai virus. J Virol. 1992 Dec;66(12):7033–7039. doi: 10.1128/jvi.66.12.7033-7039.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Pathak V. K., Temin H. M. Broad spectrum of in vivo forward mutations, hypermutations, and mutational hotspots in a retroviral shuttle vector after a single replication cycle: substitutions, frameshifts, and hypermutations. Proc Natl Acad Sci U S A. 1990 Aug;87(16):6019–6023. doi: 10.1073/pnas.87.16.6019. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Pelet T., Curran J., Kolakofsky D. The P gene of bovine parainfluenza virus 3 expresses all three reading frames from a single mRNA editing site. EMBO J. 1991 Feb;10(2):443–448. doi: 10.1002/j.1460-2075.1991.tb07966.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Rueda P., Delgado T., Portela A., Melero J. A., García-Barreno B. Premature stop codons in the G glycoprotein of human respiratory syncytial viruses resistant to neutralization by monoclonal antibodies. J Virol. 1991 Jun;65(6):3374–3378. doi: 10.1128/jvi.65.6.3374-3378.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Rueda P., García-Barreno B., Melero J. A. Loss of conserved cysteine residues in the attachment (G) glycoprotein of two human respiratory syncytial virus escape mutants that contain multiple A-G substitutions (hypermutations). Virology. 1994 Feb;198(2):653–662. doi: 10.1006/viro.1994.1077. [DOI] [PubMed] [Google Scholar]
  29. 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]
  30. Satake M., Coligan J. E., Elango N., Norrby E., Venkatesan S. Respiratory syncytial virus envelope glycoprotein (G) has a novel structure. Nucleic Acids Res. 1985 Nov 11;13(21):7795–7812. doi: 10.1093/nar/13.21.7795. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Sullender W. M., Mufson M. A., Anderson L. J., Wertz G. W. Genetic diversity of the attachment protein of subgroup B respiratory syncytial viruses. J Virol. 1991 Oct;65(10):5425–5434. doi: 10.1128/jvi.65.10.5425-5434.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Wertz G. W., Collins P. L., Huang Y., Gruber C., Levine S., Ball L. A. Nucleotide sequence of the G protein gene of human respiratory syncytial virus reveals an unusual type of viral membrane protein. Proc Natl Acad Sci U S A. 1985 Jun;82(12):4075–4079. doi: 10.1073/pnas.82.12.4075. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Wertz G. W., Krieger M., Ball L. A. Structure and cell surface maturation of the attachment glycoprotein of human respiratory syncytial virus in a cell line deficient in O glycosylation. J Virol. 1989 Nov;63(11):4767–4776. doi: 10.1128/jvi.63.11.4767-4776.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]

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