Paramecium bursaria Chlorella virus 1 encodes two enzymes involved in the biosynthesis of GDP-L-fucose and GDP-D-rhamnose

Michela Tonetti, Davide Zanardi, James R. Gurnon, Floriana Fruscione, Andrea Armirotti, Gianluca Damonte, Laura Sturla, Antonio De Flora, James L. Van Etten

Research output: Contribution to journalArticle

Abstract

At least three structural proteins in Paramecium bursaria Chlorella virus (PBCV-1) are glycosylated, including the major capsid protein Vp54. However, unlike other glycoprotein-containing viruses that use host-encoded enzymes in the endoplasmic reticulum-Golgi to glycosylate their proteins, PBCV-1 encodes at least many, if not all, of the glycosyltransferases used to glycosylate its structural proteins. As described here, PBCV-1 also encodes two open reading frames that resemble bacterial and mammalian enzymes involved in de novo GDP-L-fucose biosynthesis. This pathway, starting from GDP-D-mannose, consists of two sequential steps catalyzed by GDP-D-mannose 4,6 dehydratase (GMD) and GDP-4-keto-6-deoxy-D-mannose epimerase/reductase, respectively. The two PBCV-1-encoded genes were expressed in Escherichia coli, and the recombinant proteins had the predicted enzyme activity. However, in addition to the dehydratase activity, PBCV-1 GMD also had a reductase activity, producing GDP-D-rhamnose. In vivo studies established that PBCV-1 GMD and GDP-4-keto-6-deoxy-D-mannose epimerase/reductase are expressed after virus infection and that both GDP-L-fucose and GDP-D-rhamnose are produced in virus-infected cells. Thus, PBCV-1 is the first virus known to encode enzymes involved in nucleotide sugar metabolism. Because fucose and rhamnose are components of the glycans attached to Vp54, the pathway could circumvent a limited supply of GDP sugars by the algal host.

Original languageEnglish
Pages (from-to)21559-21565
Number of pages7
JournalJournal of Biological Chemistry
Volume278
Issue number24
DOIs
Publication statusPublished - Jun 13 2003

ASJC Scopus subject areas

  • Biochemistry

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    Tonetti, M., Zanardi, D., Gurnon, J. R., Fruscione, F., Armirotti, A., Damonte, G., Sturla, L., De Flora, A., & Van Etten, J. L. (2003). Paramecium bursaria Chlorella virus 1 encodes two enzymes involved in the biosynthesis of GDP-L-fucose and GDP-D-rhamnose. Journal of Biological Chemistry, 278(24), 21559-21565. https://doi.org/10.1074/jbc.M301543200