The N-glycan structures of the antigenic variants of chlorovirus PBCV-1 major capsid protein help to identify the virus-encoded glycosyltransferases

Immacolata Speciale, Garry A. Duncan, Luca Unione, Irina V. Agarkova, Domenico Garozzo, Jesus Jimenez-Barbero, Sicheng Lin, Todd L. Lowary, Antonio Molinaro, Eric Noel, Maria Elena Laugieri, Michela G. Tonetti, James L. Van Etten, Cristina De Castro

Research output: Contribution to journalArticle

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Abstract

The chlorovirus Paramecium bursaria chlorella virus 1 (PBCV-1) is a large dsDNA virus that infects the microalga Chlorella variabilis NC64A. Unlike most other viruses, PBCV-1 encodes most, if not all, of the machinery required to glycosylate its major capsid protein (MCP). The structures of the four N-linked glycans from the PBCV-1 MCP consist of nonasaccharides, and similar glycans are not found elsewhere in the three domains of life. Here, we identified the roles of three virus-encoded glycosyltransferases (GTs) that have four distinct GT activities in glycan synthesis. Two of the three GTs were previously annotated as GTs, but the third GT was identified in this study. We determined the GT functions by comparing the WT glycan structures from PBCV-1 with those from a set of PBCV-1 spontaneous GT gene mutants resulting in antigenic variants having truncated glycan structures. According to our working model, the virus gene a064r encodes a GT with three domains: domain 1 has α-L-rhamnosyltransferase activity, domain 2 has an β-L-rhamnosyltransferase activity, and domain 3 is a methyltransferase that decorates two positions in the terminal β-L-rhamnose (Rha) unit. The a075l gene encodes α-xylosyltransferase that attaches the distal D-xylose (Xyl) unit to the L-fucose (Fuc) that is part of the conserved N-glycan core region. Last, gene a071r encodes a GT that is involved in the attachment of a semiconserved element, β-D-Rha, to the same L-Fuc in the core region. Our results uncover GT activities that assemble four of the nine residues of the PBCV-1 MCP N-glycans.

Original languageEnglish (US)
Pages (from-to)5688-5699
Number of pages12
JournalJournal of Biological Chemistry
Volume294
Issue number14
DOIs
StatePublished - Jan 1 2019

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Paramecium
Chlorella
Glycosyltransferases
Capsid Proteins
Viruses
Polysaccharides
Genes
Fucose
Rhamnose
Xylose
Methyltransferases
Machinery

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

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The N-glycan structures of the antigenic variants of chlorovirus PBCV-1 major capsid protein help to identify the virus-encoded glycosyltransferases. / Speciale, Immacolata; Duncan, Garry A.; Unione, Luca; Agarkova, Irina V.; Garozzo, Domenico; Jimenez-Barbero, Jesus; Lin, Sicheng; Lowary, Todd L.; Molinaro, Antonio; Noel, Eric; Laugieri, Maria Elena; Tonetti, Michela G.; Van Etten, James L.; De Castro, Cristina.

In: Journal of Biological Chemistry, Vol. 294, No. 14, 01.01.2019, p. 5688-5699.

Research output: Contribution to journalArticle

Speciale, I, Duncan, GA, Unione, L, Agarkova, IV, Garozzo, D, Jimenez-Barbero, J, Lin, S, Lowary, TL, Molinaro, A, Noel, E, Laugieri, ME, Tonetti, MG, Van Etten, JL & De Castro, C 2019, 'The N-glycan structures of the antigenic variants of chlorovirus PBCV-1 major capsid protein help to identify the virus-encoded glycosyltransferases', Journal of Biological Chemistry, vol. 294, no. 14, pp. 5688-5699. https://doi.org/10.1074/jbc.RA118.007182
Speciale, Immacolata ; Duncan, Garry A. ; Unione, Luca ; Agarkova, Irina V. ; Garozzo, Domenico ; Jimenez-Barbero, Jesus ; Lin, Sicheng ; Lowary, Todd L. ; Molinaro, Antonio ; Noel, Eric ; Laugieri, Maria Elena ; Tonetti, Michela G. ; Van Etten, James L. ; De Castro, Cristina. / The N-glycan structures of the antigenic variants of chlorovirus PBCV-1 major capsid protein help to identify the virus-encoded glycosyltransferases. In: Journal of Biological Chemistry. 2019 ; Vol. 294, No. 14. pp. 5688-5699.
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abstract = "The chlorovirus Paramecium bursaria chlorella virus 1 (PBCV-1) is a large dsDNA virus that infects the microalga Chlorella variabilis NC64A. Unlike most other viruses, PBCV-1 encodes most, if not all, of the machinery required to glycosylate its major capsid protein (MCP). The structures of the four N-linked glycans from the PBCV-1 MCP consist of nonasaccharides, and similar glycans are not found elsewhere in the three domains of life. Here, we identified the roles of three virus-encoded glycosyltransferases (GTs) that have four distinct GT activities in glycan synthesis. Two of the three GTs were previously annotated as GTs, but the third GT was identified in this study. We determined the GT functions by comparing the WT glycan structures from PBCV-1 with those from a set of PBCV-1 spontaneous GT gene mutants resulting in antigenic variants having truncated glycan structures. According to our working model, the virus gene a064r encodes a GT with three domains: domain 1 has α-L-rhamnosyltransferase activity, domain 2 has an β-L-rhamnosyltransferase activity, and domain 3 is a methyltransferase that decorates two positions in the terminal β-L-rhamnose (Rha) unit. The a075l gene encodes α-xylosyltransferase that attaches the distal D-xylose (Xyl) unit to the L-fucose (Fuc) that is part of the conserved N-glycan core region. Last, gene a071r encodes a GT that is involved in the attachment of a semiconserved element, β-D-Rha, to the same L-Fuc in the core region. Our results uncover GT activities that assemble four of the nine residues of the PBCV-1 MCP N-glycans.",
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AU - Duncan, Garry A.

AU - Unione, Luca

AU - Agarkova, Irina V.

AU - Garozzo, Domenico

AU - Jimenez-Barbero, Jesus

AU - Lin, Sicheng

AU - Lowary, Todd L.

AU - Molinaro, Antonio

AU - Noel, Eric

AU - Laugieri, Maria Elena

AU - Tonetti, Michela G.

AU - Van Etten, James L.

AU - De Castro, Cristina

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