Grapevine fanleaf virus replication occurs on endoplasmic reticulum-derived membranes.

  • Ritzenthaler C
  • Laporte C
  • Gaire F
 et al. 
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Abstract

Infection by Grapevine fanleaf nepovirus (GFLV), a bipartite RNA virus of positive polarity belonging to the Comoviridae family, causes extensive cytopathic modifications of the host endomembrane system that eventually culminate in the formation of a perinuclear "viral compartment." We identified by immunoconfocal microscopy this compartment as the site of virus replication since it contained the RNA1-encoded proteins necessary for replication, newly synthesized viral RNA, and double-stranded replicative forms. In addition, by using transgenic T-BY2 protoplasts expressing green fluorescent protein in the endoplasmic reticulum (ER) or in the Golgi apparatus (GA), we could directly show that GFLV replication induced a depletion of the cortical ER, together with a condensation and redistribution of ER-derived membranes, to generate the viral compartment. Brefeldin A, a drug known to inhibit vesicle trafficking between the GA and the ER, was found to inhibit GFLV replication. Cerulenin, a drug inhibiting de novo synthesis of phospholipids, also inhibited GFLV replication. These observations imply that GFLV replication depends both on ER-derived membrane recruitment and on de novo lipid synthesis. In contrast to proteins involved in viral replication, the 2B movement protein and, to a lesser extent, the 2C coat protein were not confined to the viral compartment but were transported toward the cell periphery, a finding consistent with their role in cell-to-cell movement of virus particles.

Author-supplied keywords

  • 00 ϩ 0 doi
  • 0022-538x
  • 02
  • 04
  • 10
  • 1128
  • 17
  • 2002
  • 76
  • 8808
  • 8819
  • all rights reserved
  • american society for microbiology
  • copyright
  • jvi
  • p
  • sept
  • urnal of v irology

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Authors

  • C Ritzenthaler

  • C Laporte

  • F Gaire

  • P Dunoyer

  • C Schmitt

  • S Duval

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