title: Cumulative mutations in the genome of Echovirus 6 during establishment of a chronic infection in precursors of glial cells.
abstract: Although Enteroviruses are mainly described as responsible for acute diseases, their role in severe chronic pathology has been also established. Echovirus 6-like sequences have been detected by PCR analysis in central nervous system specimens from patients presenting with Amyotrophic Lateral Sclerosis. These findings suggested a persistent infection with viruses that underwent, genetic changes precluding viral particle release. To support this hypothesis, we developed a model system of Echovirus 6 chronic infection in precursors of glial cells. The nucleotide sequences of the 5'non-translated region (5'NTR), 2A and 3C regions of the virus developing persistent genome were analysed during establishment of the chronic phenotype. This study revealed that at day 160 of chronic infection, several mutations were observed: one mutation at nucleotide 108 upstream the domain II of the internal ribosome entry site (IRES) structure, one mutation at nucleotide 30 in the cloverleaf, and two mutations in the 2A region (translated in His48 to Tyr, and Ile 123 to Met). No mutations were detected in the 3C region. The impact of these mutations on viral replication have been analysed in a rabbit reticulocyte lysate (RRL) translation assay supplemented with HeLa cell lysate, and by plaque assay. Viruses with these mutations displayed a phenotype with a significant reduction of replication, while in vitro translation was not affected by the nucleotide 108 mutation. This model allowed the description of molecular changes observed in the genome of Echovirus 6 during the establishment of a chronic infection phenotype, and may be helpful for the understanding of the mechanisms leading Enteroviruses to develop chronic infections in man.
authors.affiliation: Laboratoire de Virologie, UMR CNRS 5537, Domaine Rockefeller, F-69373, Lyon cedex 08, France.
authors.forename: Frederik
authors.initials: F
authors.lastname: Beaulieux
authors.forename: Youssef
authors.initials: Y
authors.lastname: Zreik
authors.forename: Christelle
authors.initials: C
authors.lastname: Deleage
authors.forename: Valerie
authors.initials: V
authors.lastname: Sauvinet
authors.forename: Vincent
authors.initials: V
authors.lastname: Legay
authors.forename: Pascale
authors.initials: P
authors.lastname: Giraudon
authors.forename: Katherine M
authors.initials: KM
authors.lastname: Kean
authors.forename: Bruno
authors.initials: B
authors.lastname: Lina
mesh_terms: D020121:5' Untranslated Regions; D000595:Amino Acid Sequence; D019943:Amino Acid Substitution; D000690:Amyotrophic Lateral Sclerosis / Q000821:virology; D001483:Base Sequence; D002460:Cell Line; D004458:Echovirus 6, Human / Q000235:genetics* / Q000502:physiology*; D004457:Echovirus Infections / Q000821:virology; D006801:Humans; D009154:Mutation; D020125:Mutation, Missense; D009457:Neuroglia / Q000821:virology*; D009690:Nucleic Acid Conformation; D017354:Point Mutation; D012367:RNA, Viral / Q000235:genetics; D017421:Sequence Analysis; D010948:Viral Plaque Assay; D014764:Viral Proteins / Q000235:genetics; D014779:Virus Replication
chemical_list: D020121:5' Untranslated Regions; D012367:RNA, Viral; D014764:Viral Proteins
references.citation: J Virol. 1995 Jun;69(6):3465-74
references.pmid: 7745693
references.citation: J Virol. 2001 Nov;75(22):10643-50
references.pmid: 11602706
references.citation: Med Microbiol Immunol. 2004 May;193(2-3):121-6
references.pmid: 12920582
references.citation: J Virol. 1985 Apr;54(1):219-23
references.pmid: 3973978
references.citation: Proc Natl Acad Sci U S A. 1989 Oct;86(19):7590-4
references.pmid: 2552448
references.citation: Viral Immunol. 1999;12(4):335-41
references.pmid: 10630792
references.citation: Proc Natl Acad Sci U S A. 1992 Jan 1;89(1):314-8
references.pmid: 1309611
references.citation: Eur J Neurosci. 2003 May;17(9):1820-8
references.pmid: 12752781
references.citation: Nucleic Acids Res. 1989 Apr 25;17(8):3129-44
references.pmid: 2726454
references.citation: J Virol. 2000 Mar;74(5):2219-26
references.pmid: 10666252
references.citation: Cell. 1992 Jan 10;68(1):119-31
references.pmid: 1310072
references.citation: FEBS Lett. 2002 Jul 17;523(1-3):53-7
references.pmid: 12123803
references.citation: EMBO J. 1994 Jul 1;13(13):3149-57
references.pmid: 8039507
references.citation: Proc Natl Acad Sci U S A. 1994 Apr 26;91(9):3705-9
references.pmid: 8170973
references.citation: J Biol Chem. 2000 Oct 13;275(41):32268-76
references.pmid: 10922367
references.citation: Nucleic Acids Res. 1995 Sep 25;23(18):3656-63
references.pmid: 7478993
references.citation: J Virol. 1989 Dec;63(12):5268-75
references.pmid: 2585604
references.citation: J Virol. 2000 Oct;74(19):9071-7
references.pmid: 10982352
references.citation: Virology. 2000 Apr 10;269(2):284-93
references.pmid: 10753707
references.citation: J Infect Dis. 1997 Mar;175(3):693-6
references.pmid: 9041346
references.citation: J Virol. 1993 May;67(5):2914-7
references.pmid: 8386281
references.citation: J Virol. 1990 Oct;64(10):4922-9
references.pmid: 2168976
references.citation: Trends Microbiol. 2000 Jul;8(7):330-5
references.pmid: 10878768
references.citation: Neuron. 1998 Mar;20(3):589-602
references.pmid: 9539131
references.citation: Genes Dev. 1998 Aug 1;12(15):2293-304
references.pmid: 9694795
references.citation: Virology. 1996 Jan 1;215(1):83-96
references.pmid: 8553589
references.citation: Neurology. 2000 Jan 11;54(1):20-5
references.pmid: 10636120
references.citation: J Virol. 1995 Jan;69(1):247-52
references.pmid: 7983716
references.citation: J Med Virol. 1997 Jun;52(2):121-7
references.pmid: 9179756
references.citation: Mol Biol Rep. 1994 May;19(3):147-59
references.pmid: 7969103
references.citation: Neurology. 2001 Jun 26;56(12):1777-8
references.pmid: 11425957
references.citation: J Gen Virol. 2000 Nov;81(Pt 11):2755-62
references.pmid: 11038389
references.citation: Rev Neurol (Paris). 2000 Apr;156(4):352-6
references.pmid: 10795011
references.citation: J Gen Virol. 1990 Jun;71 ( Pt 6):1399-402
references.pmid: 2161907
references.citation: Mol Cell Probes. 1994 Dec;8(6):487-95
references.pmid: 7700271
references.citation: Eur J Cancer. 1992;28A(4-5):762-6
references.pmid: 1326305
references.citation: J Virol. 2001 Jul;75(14):6472-81
references.pmid: 11413314