Characterization and construction of functional cDNA clones of pariacoto virus, the first Alphanodavirus isolated outside Australasia

Johnson, Karyn N., Zeddam, Jean-Louis and Ball, L. Andrew (2000) Characterization and construction of functional cDNA clones of pariacoto virus, the first Alphanodavirus isolated outside Australasia. Journal of Virology, 74 11: 5123-5132. doi:10.1128/JVI.74.11.5123-5132.2000


Author Johnson, Karyn N.
Zeddam, Jean-Louis
Ball, L. Andrew
Title Characterization and construction of functional cDNA clones of pariacoto virus, the first Alphanodavirus isolated outside Australasia
Journal name Journal of Virology   Check publisher's open access policy
ISSN 0022-538X
1098-5514
Publication date 2000-06-01
Year available 2000
Sub-type Article (original research)
DOI 10.1128/JVI.74.11.5123-5132.2000
Open Access Status DOI
Volume 74
Issue 11
Start page 5123
End page 5132
Total pages 10
Place of publication Washington, DC, United States
Publisher American Society for Microbiology
Language eng
Formatted abstract
Pariacoto virus (PaV) was recently isolated in Peru from the Southern armyworm (Spodoptera eridania). PaV particles are isometric, nonenveloped, and about 30 nm in diameter. The virus has a bipartite RNA genome and a single major capsid protein with a molecular mass of 39.0 kDa, features that support its classification as aNodavirus. As such, PaV is the firstAlphanodavirus to have been isolated from outside Australasia. Here we report that PaV replicates in wax moth larvae and that PaV genomic RNAs replicate when transfected into cultured baby hamster kidney cells. The complete nucleotide sequences of both segments of the bipartite RNA genome were determined. The larger genome segment, RNA1, is 3,011 nucleotides long and contains a 973-amino-acid open reading frame (ORF) encoding protein A, the viral contribution to the RNA replicase. During replication, a 414-nucleotide long subgenomic RNA (RNA3) is synthesized which is coterminal with the 3′ end of RNA1. RNA3 contains a small ORF which could encode a protein of 90 amino acids similar to the B2 protein of other alphanodaviruses. RNA2 contains 1,311 nucleotides and encodes the 401 amino acids of the capsid protein precursor α. The amino acid sequences of the PaV capsid protein and the replicase subunit share 41 and 26% identity with homologous proteins of Flock house virus, the best characterized of the alphanodaviruses. These and other sequence comparisons indicate that PaV is evolutionarily the most distant of the alphanodaviruses described to date, consistent with its novel geographic origin. Although the PaV capsid precursor is cleaved into the two mature capsid proteins β and γ, the amino acid sequence at the cleavage site, which is Asn/Ala in all other alphanodaviruses, is Asn/Ser in PaV. To facilitate the investigation of PaV replication in cultured cells, we constructed plasmids that transcribed full-length PaV RNAs with authentic 5′ and 3′ termini. Transcription of these plasmids in cells recreated the replication of PaV RNA1 and RNA2, synthesis of subgenomic RNA3, and translation of viral proteins A and α.
Keyword Black Beetle Virus
Flock House Virus
Insect Virus
Genomic Rna
Nervous Necrosis
Fish Nodaviruses
Vaccinia Virus
Animal Virus
Protein Gene
Replication
Q-Index Code C1
Q-Index Status Provisional Code
Institutional Status Unknown

Document type: Journal Article
Sub-type: Article (original research)
Collection: ResearcherID Downloads
 
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