Synthesis of infectious transcripts of olive latent virus 1: genes required for RNA replication and virus movement

Synthesis of infectious transcripts of olive latent virus 1: genes required for RNA replication... Genomic RNA of olive latent virus 1 (OLV-1) contains five open reading frames (ORFs) encoding proteins of 23, 82, 8, 6 and 30 (CP) kDa. A full-length cDNA copy of OLV-1 RNA was prepared and cloned in a low-copy-number vector (pMUC-19) downstream of or T7 RNA polymerase promoter. Transcripts derived from this template, denoted pMUC-OLV, were highly infectious when inoculated in local and systemic hosts and infected tissues contained virus-like particles. Genes required for replication and virus movement were mapped by site-directed and deletion mutogenesis of the pMUC-OLV. ORF1 and ORF2 mutants were not viable, suggesting that replication requires the 23 and 82 kDa proteins. The 8 and 6 kDa polypeptides were involved in cell-to-cell movement, since their absence did not interfere with RNA replication but prevented systemic infection of inoculated plants. Mutant clones in R and S domains of the CP gene could replicate, but they did not systemically infect Nicotiana benthamiana , indicating that the CP gene is required for OLV-1 long-distance translocation. Mutant clones with large deletions in the CP gene were not viable, probably due to loss of 3′-proximal sequences required for RNA replication. http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png Archives of Virology Springer Journals

Synthesis of infectious transcripts of olive latent virus 1: genes required for RNA replication and virus movement

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Publisher
Springer-Verlag
Copyright
Copyright © Wien by 1999 Springer-Verlag/
Subject
Legacy
ISSN
0304-8608
eISSN
1432-8798
D.O.I.
10.1007/s007050050570
Publisher site
See Article on Publisher Site

Abstract

Genomic RNA of olive latent virus 1 (OLV-1) contains five open reading frames (ORFs) encoding proteins of 23, 82, 8, 6 and 30 (CP) kDa. A full-length cDNA copy of OLV-1 RNA was prepared and cloned in a low-copy-number vector (pMUC-19) downstream of or T7 RNA polymerase promoter. Transcripts derived from this template, denoted pMUC-OLV, were highly infectious when inoculated in local and systemic hosts and infected tissues contained virus-like particles. Genes required for replication and virus movement were mapped by site-directed and deletion mutogenesis of the pMUC-OLV. ORF1 and ORF2 mutants were not viable, suggesting that replication requires the 23 and 82 kDa proteins. The 8 and 6 kDa polypeptides were involved in cell-to-cell movement, since their absence did not interfere with RNA replication but prevented systemic infection of inoculated plants. Mutant clones in R and S domains of the CP gene could replicate, but they did not systemically infect Nicotiana benthamiana , indicating that the CP gene is required for OLV-1 long-distance translocation. Mutant clones with large deletions in the CP gene were not viable, probably due to loss of 3′-proximal sequences required for RNA replication.

Journal

Archives of VirologySpringer Journals

Published: Jun 1, 1999

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