Isolation of enzymatically active replication complexes from feline calicivirus-infected cells

Kim Y. Green, Aaron Mory, Mark H. Fogg, Andrea Weisberg, Gaël Belliot, Mariam Wagner, Tanaji Mitra, Ellie Ehrenfeld, Craig E. Cameron, Stanislav V. Sosnovtsev

Research output: Contribution to journalArticle

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Abstract

A membranous fraction that could synthesize viral RNA in vitro in the presence of magnesium salt, ribonucleotides, and an ATP-regenerating system was isolated from feline calicivirus (FCV)-infected cells. The enzymatically active component of this fraction was designated FCV replication complexes (RCs), by analogy to other positive-strand RNA viruses. The newly synthesized RNA was characterized by Northern blot analysis, which demonstrated the production of both full-length (8.0-kb) and subgenomic-length (2.5-kb) RNA molecules similar to those synthesized in FCV-infected cells. The identity of the viral proteins associated with the fraction was investigated. The 60-kDa VP1 major capsid protein was the most abundant viral protein detected. VP2, a minor structural protein encoded by open reading frame 3 (ORF3), was also present. Nonstructural proteins associated with the fraction included the precursor polypeptides Pro-Pol (76 kDa) and p30-VPg (43 kDa), as well as the mature nonstructural proteins p32 (derived from the N-terminal region of the ORF1 polyprotein), p30 (the putative "3A-like" protein), and p39 (the putative nucleoside triphosphatase). The isolation of enzymatically active RCs containing both viral and cellular proteins should facilitate efforts to dissect the contributions of the virus and the host to FCV RNA replication.

Original languageEnglish (US)
Pages (from-to)8582-8595
Number of pages14
JournalJournal of virology
Volume76
Issue number17
DOIs
StatePublished - Aug 20 2002

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Feline Calicivirus
Feline calicivirus
Viral Proteins
viral proteins
RNA
Proteins
proteins
Nucleoside-Triphosphatase
cells
ribonucleotides
Ribonucleotides
Polyproteins
RNA Viruses
structural proteins
Viral RNA
Capsid Proteins
coat proteins
Northern blotting
Northern Blotting
Magnesium

All Science Journal Classification (ASJC) codes

  • Microbiology
  • Immunology
  • Insect Science
  • Virology

Cite this

Green, K. Y., Mory, A., Fogg, M. H., Weisberg, A., Belliot, G., Wagner, M., ... Sosnovtsev, S. V. (2002). Isolation of enzymatically active replication complexes from feline calicivirus-infected cells. Journal of virology, 76(17), 8582-8595. https://doi.org/10.1128/JVI.76.17.8582-8595.2002
Green, Kim Y. ; Mory, Aaron ; Fogg, Mark H. ; Weisberg, Andrea ; Belliot, Gaël ; Wagner, Mariam ; Mitra, Tanaji ; Ehrenfeld, Ellie ; Cameron, Craig E. ; Sosnovtsev, Stanislav V. / Isolation of enzymatically active replication complexes from feline calicivirus-infected cells. In: Journal of virology. 2002 ; Vol. 76, No. 17. pp. 8582-8595.
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Green, KY, Mory, A, Fogg, MH, Weisberg, A, Belliot, G, Wagner, M, Mitra, T, Ehrenfeld, E, Cameron, CE & Sosnovtsev, SV 2002, 'Isolation of enzymatically active replication complexes from feline calicivirus-infected cells', Journal of virology, vol. 76, no. 17, pp. 8582-8595. https://doi.org/10.1128/JVI.76.17.8582-8595.2002

Isolation of enzymatically active replication complexes from feline calicivirus-infected cells. / Green, Kim Y.; Mory, Aaron; Fogg, Mark H.; Weisberg, Andrea; Belliot, Gaël; Wagner, Mariam; Mitra, Tanaji; Ehrenfeld, Ellie; Cameron, Craig E.; Sosnovtsev, Stanislav V.

In: Journal of virology, Vol. 76, No. 17, 20.08.2002, p. 8582-8595.

Research output: Contribution to journalArticle

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T1 - Isolation of enzymatically active replication complexes from feline calicivirus-infected cells

AU - Green, Kim Y.

AU - Mory, Aaron

AU - Fogg, Mark H.

AU - Weisberg, Andrea

AU - Belliot, Gaël

AU - Wagner, Mariam

AU - Mitra, Tanaji

AU - Ehrenfeld, Ellie

AU - Cameron, Craig E.

AU - Sosnovtsev, Stanislav V.

PY - 2002/8/20

Y1 - 2002/8/20

N2 - A membranous fraction that could synthesize viral RNA in vitro in the presence of magnesium salt, ribonucleotides, and an ATP-regenerating system was isolated from feline calicivirus (FCV)-infected cells. The enzymatically active component of this fraction was designated FCV replication complexes (RCs), by analogy to other positive-strand RNA viruses. The newly synthesized RNA was characterized by Northern blot analysis, which demonstrated the production of both full-length (8.0-kb) and subgenomic-length (2.5-kb) RNA molecules similar to those synthesized in FCV-infected cells. The identity of the viral proteins associated with the fraction was investigated. The 60-kDa VP1 major capsid protein was the most abundant viral protein detected. VP2, a minor structural protein encoded by open reading frame 3 (ORF3), was also present. Nonstructural proteins associated with the fraction included the precursor polypeptides Pro-Pol (76 kDa) and p30-VPg (43 kDa), as well as the mature nonstructural proteins p32 (derived from the N-terminal region of the ORF1 polyprotein), p30 (the putative "3A-like" protein), and p39 (the putative nucleoside triphosphatase). The isolation of enzymatically active RCs containing both viral and cellular proteins should facilitate efforts to dissect the contributions of the virus and the host to FCV RNA replication.

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