Functional characterization of the alphavirus TF protein

Jonathan E. Snyder, Kirsten A. Kulcsar, Kimberly L.W. Schultz, Catherine P. Riley, Jacob T. Neary, Scott Marr, Joyce Jose, Diane E. Griffin, Richard R. Kuhn

Research output: Contribution to journalArticle

45 Citations (Scopus)

Abstract

Al phavirus dogma has long dictated the production of a discrete set of structural proteins during infection of a cell: capsid, pE2, 6K, and E1. However, bioinformatic analyses of alphavirus genomes (A. E. Firth, B. Y. Chung, M. N. Fleeton, and J. F. Atkins, Virol. J. 5:108, 2008) suggested that a ribosomal frameshifting event occurs during translation of the alphavirus structural polyprotein. Specifically, a frameshift event is suggested to occur during translation of the 6K gene, yielding production of a novel protein, termed transframe (TF), comprised of a C-terminal extension of the 6K protein in the-1 open reading frame (ORF). Here, we validate the findings of Firth and colleagues with respect to the production of the TF protein and begin to characterize the function of TF. Using a mass spectrometry-based approach, we identified TF in purified preparations of both Sindbis and Chikungunya virus particles. We next constructed a panel of Sindbis virus mutants with mutations which alter the production, size, or sequence of TF. We demonstrate that TF is not absolutely required in culture, although disrupting TF production leads to a decrease in virus particle release in both mammalian and insect cells. In a mouse neuropathogenesis model, mortality was <15% in animals infected with the TF mutants, whereas mortality was 95% in animals infected with the wild-type virus. Using a variety of additional assays, we demonstrate that TF retains ion-channel activity analogous to that of 6K and that lack of production of TF does not affect genome replication, particle infectivity, or envelope protein transit to the cell surface. The TF protein therefore represents a previously uncharacterized factor important for alphavirus assembly.

Original languageEnglish (US)
Pages (from-to)8511-8523
Number of pages13
JournalJournal of Virology
Volume87
Issue number15
DOIs
StatePublished - Aug 2 2013

Fingerprint

Alphavirus
Sindbis virus
Sindbis Virus
Proteins
virion
proteins
translation (genetics)
Virion
Chikungunya virus
Ribosomal Frameshifting
mutants
Genome
capsid
genome
structural proteins
Virus Release
ion channels
Polyproteins
cells
bioinformatics

All Science Journal Classification (ASJC) codes

  • Microbiology
  • Immunology
  • Insect Science
  • Virology

Cite this

Snyder, J. E., Kulcsar, K. A., Schultz, K. L. W., Riley, C. P., Neary, J. T., Marr, S., ... Kuhn, R. R. (2013). Functional characterization of the alphavirus TF protein. Journal of Virology, 87(15), 8511-8523. https://doi.org/10.1128/JVI.00449-13
Snyder, Jonathan E. ; Kulcsar, Kirsten A. ; Schultz, Kimberly L.W. ; Riley, Catherine P. ; Neary, Jacob T. ; Marr, Scott ; Jose, Joyce ; Griffin, Diane E. ; Kuhn, Richard R. / Functional characterization of the alphavirus TF protein. In: Journal of Virology. 2013 ; Vol. 87, No. 15. pp. 8511-8523.
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Snyder, JE, Kulcsar, KA, Schultz, KLW, Riley, CP, Neary, JT, Marr, S, Jose, J, Griffin, DE & Kuhn, RR 2013, 'Functional characterization of the alphavirus TF protein', Journal of Virology, vol. 87, no. 15, pp. 8511-8523. https://doi.org/10.1128/JVI.00449-13

Functional characterization of the alphavirus TF protein. / Snyder, Jonathan E.; Kulcsar, Kirsten A.; Schultz, Kimberly L.W.; Riley, Catherine P.; Neary, Jacob T.; Marr, Scott; Jose, Joyce; Griffin, Diane E.; Kuhn, Richard R.

In: Journal of Virology, Vol. 87, No. 15, 02.08.2013, p. 8511-8523.

Research output: Contribution to journalArticle

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AU - Kuhn, Richard R.

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Snyder JE, Kulcsar KA, Schultz KLW, Riley CP, Neary JT, Marr S et al. Functional characterization of the alphavirus TF protein. Journal of Virology. 2013 Aug 2;87(15):8511-8523. https://doi.org/10.1128/JVI.00449-13