The cardiovirulent phenotype of coxsackievirus B3 is determined at a single site in the genomic 5′ nontranslated region

Zhongheng Tu, Nora M Chapman, Günter Hufnagel, Steven M Tracy, José R. Romero, William H. Barry, Liping Zhao, Kathleen Currey, Bruce Shapiro

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Abstract

We report the construction of chimeric coxsackievirus B3 (CVB3) strains in which sequences of an infectious cDNA copy of a noncardiovirulent CVB3 genome were replaced by the homologous sequences from a cardiovirulent CVB3 genome to identify which of 10 predicted genetic sites determine cardiovirulence. Cardiovirulent phenotype expression was consistently linked to nucleotide 234 (U in cardiovirulent CVB3 and C in avirulent CVB3) in the 5′ nontranslated region. Reconstructions of the parental noncardiovirulent CVB3 genome from chimeras restored the noncardiovirulent phenotype when tested in mice. Inoculation of severe combined immunodeficient (scid) mice with the noncardiovirulent CVB3 strain resulted in massive cardiomyocyte necrosis in all animals. Sequence analysis of viral genomes isolated from twelve scid mouse hearts showed that only nucleotide position 234 was different (a C→U transition) from that in the input parental noncardiovirulent CVB3 genome. Higher-order RNA structures predicted by two different algorithms did not demonstrate an obvious local effect caused by the C→U change at nucleotide 234. Initial studies of parental and chimeric CVB3 replication in primary cultures of fetal murine heart fibroblasts and in adult murine cardiac myocytes demonstrated that viral RNA transcriptional efficiency is approximately 10-fold lower for noncardiovirulent CVB3 than for cardiovirulent CVB3. CVB3 did not shut off protein synthesis in murine cardiac fibroblasts, nor were levels of viral protein synthesis significantly different as a function of viral phenotype. Taken together, these data support a significant role for determination of the CVB3 cardiovirulence phenotype by nucleotide 234 in the 5′ nontranslated region, possibly via a transcriptional mechanism.

Original languageEnglish (US)
Pages (from-to)4607-4618
Number of pages12
JournalJournal of virology
Volume69
Issue number8
StatePublished - Aug 1 1995

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Enterovirus
Phenotype
genomics
phenotype
mice
Nucleotides
nucleotides
genome
Genome
SCID Mice
Cardiac Myocytes
fibroblasts
Fibroblasts
protein synthesis
heart
RNA
Fetal Heart
chimerism
Viral Genome
viral proteins

ASJC Scopus subject areas

  • Immunology

Cite this

Tu, Z., Chapman, N. M., Hufnagel, G., Tracy, S. M., Romero, J. R., Barry, W. H., ... Shapiro, B. (1995). The cardiovirulent phenotype of coxsackievirus B3 is determined at a single site in the genomic 5′ nontranslated region. Journal of virology, 69(8), 4607-4618.

The cardiovirulent phenotype of coxsackievirus B3 is determined at a single site in the genomic 5′ nontranslated region. / Tu, Zhongheng; Chapman, Nora M; Hufnagel, Günter; Tracy, Steven M; Romero, José R.; Barry, William H.; Zhao, Liping; Currey, Kathleen; Shapiro, Bruce.

In: Journal of virology, Vol. 69, No. 8, 01.08.1995, p. 4607-4618.

Research output: Contribution to journalArticle

Tu, Z, Chapman, NM, Hufnagel, G, Tracy, SM, Romero, JR, Barry, WH, Zhao, L, Currey, K & Shapiro, B 1995, 'The cardiovirulent phenotype of coxsackievirus B3 is determined at a single site in the genomic 5′ nontranslated region', Journal of virology, vol. 69, no. 8, pp. 4607-4618.
Tu Z, Chapman NM, Hufnagel G, Tracy SM, Romero JR, Barry WH et al. The cardiovirulent phenotype of coxsackievirus B3 is determined at a single site in the genomic 5′ nontranslated region. Journal of virology. 1995 Aug 1;69(8):4607-4618.
Tu, Zhongheng ; Chapman, Nora M ; Hufnagel, Günter ; Tracy, Steven M ; Romero, José R. ; Barry, William H. ; Zhao, Liping ; Currey, Kathleen ; Shapiro, Bruce. / The cardiovirulent phenotype of coxsackievirus B3 is determined at a single site in the genomic 5′ nontranslated region. In: Journal of virology. 1995 ; Vol. 69, No. 8. pp. 4607-4618.
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