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Mutational analysis of the proteinase function of Potato leafroll virus.
- Source :
-
The Journal of general virology [J Gen Virol] 2001 Jun; Vol. 82 (Pt 6), pp. 1517-1527. - Publication Year :
- 2001
-
Abstract
- cDNA expression vectors of Potato leafroll virus (PLRV) were used to analyse specific mutations in the proteinase and replicase domains of the proteins encoded by ORF1 and ORF2. Agrobacterium-mediated DNA transfer was used to introduce a PLRV RNA expression unit, controlled by the 35S promoter of Cauliflower mosaic virus, into potato leaf cells. Expression of unmodified PLRV cDNA led to the replication of viral genomic and subgenomic RNAs and accumulation of the viral capsid protein, whereas alteration of amino acids GDD513-515 of the replicase to VHD abolished PLRV replication. Mutations in the presumed H-D-S catalytic triad of the viral proteinase abolished the formation of viral genomic and subgenomic RNAs as well as synthesis of the viral capsid protein. Co-agroinoculation of the GDD mutant along with any of the proteinase mutants restored virus replication in leaf discs, showing that these mutants are able to complement each other. Moreover, mutation of the postulated serine residue of the catalytic triad of the proteinase altered the pattern of proteins synthesized in vitro in comparison to wild-type, further supporting the relevance of the H-D-S motif.
- Subjects :
- Amino Acid Motifs
Amino Acid Sequence
Amino Acid Substitution genetics
Base Sequence
Capsid biosynthesis
Capsid genetics
Caulimovirus genetics
Endopeptidases chemistry
Endopeptidases genetics
Genetic Complementation Test
Genome, Viral
Luteovirus genetics
Luteovirus physiology
Open Reading Frames genetics
Plant Leaves microbiology
Plant Leaves virology
Promoter Regions, Genetic genetics
Protein Structure, Tertiary
RNA, Viral biosynthesis
RNA, Viral genetics
RNA, Viral metabolism
RNA-Dependent RNA Polymerase chemistry
RNA-Dependent RNA Polymerase genetics
RNA-Dependent RNA Polymerase metabolism
Rhizobium genetics
Serine genetics
Serine metabolism
Solanum tuberosum microbiology
Virus Replication
Endopeptidases metabolism
Luteovirus enzymology
Mutation genetics
Solanum tuberosum virology
Subjects
Details
- Language :
- English
- ISSN :
- 0022-1317
- Volume :
- 82
- Issue :
- Pt 6
- Database :
- MEDLINE
- Journal :
- The Journal of general virology
- Publication Type :
- Academic Journal
- Accession number :
- 11369899
- Full Text :
- https://doi.org/10.1099/0022-1317-82-6-1517