Assembly of turnip yellow mosaic virus replication complexes: Interaction between the proteinase and polymerase domains of the replication proteins

Jakubiec, A., Notaise, J., Tournier, V., Hericourt, F., Block, M. A., Drugeon, G., van Aelst, L., Jupin, I. (August 2004) Assembly of turnip yellow mosaic virus replication complexes: Interaction between the proteinase and polymerase domains of the replication proteins. Journal of Virology, 78 (15). pp. 7945-7957. ISSN 0022-538X

DOI: 10.1128/JVI.78.15.7945-7957.2004


Turnip yellow mosaic virus (TYMV), a positive-strand RNA virus in the alphavirus-like supergroup, encodes two nonstructural replication proteins (140K and 66K), both of which are required for its RNA genome replication. The 140K protein contains domains indicative of methyltransferase, proteinase, and NTPase/helicase activities, while the 66K protein encompasses the RNA-dependent RNA polymerase domain. Recruitment of the 66K protein to the sites of viral replication, located at the periphery of chloroplasts, is dependent upon the expression of the 140K protein. Using antibodies raised against the 140K and 66K proteins and confocal microscopy, we report the colocalization of the TYMV replication proteins at the periphery of chloroplasts in transfected or infected cells. The replication proteins cofractionated in functional replication complexes or with purified chloroplast envelope membranes prepared from infected plants. Using a two-hybrid system and coimmunoprecipitation experiments, we also provide evidence for a physical interaction of the TYMV replication proteins. In contrast to what has been found for other members of the alphavirus-like supergroup, the interaction domains were mapped to the proteinase domain of the 140K protein and to a large region encompassing the core polymerase domain within the 66K protein. Coexpression and colocalization experiments confirmed that the helicase domain of the 140K protein is unnecessary for the proper recruitment of the 66K protein to the chloroplast envelope, while the proteinase domain appears to be essential for that process. These results support a novel model for the interaction of TYMV replication proteins and suggest that viruses in the alphavirus-like supergroup may have selected different pathways to assemble their replication complexes.

Item Type: Paper
Uncontrolled Keywords: STRAND RNA VIRUSES Strand RNA virus TOBACCO ETCH POTYVIRUS Tobacco Etch Potyvirus VIRAL PROTEIN viral protein ENDOPLASMIC-RETICULUM endoplasmic reticulum FUNCTIONAL-ANALYSIS functional analysis ESCHERICHIA-COLI escherichia coli 2-HYBRID 2-Hybrid SYSTEM CLEAVAGE SITE cleavage site HELICASE-LIKE helicase like GENOMIC RNA genomic RNA
Subjects: bioinformatics > genomics and proteomics > genetics & nucleic acid processing > DNA, RNA structure, function, modification > RNA replication
organism description > virus
CSHL Authors:
Communities: CSHL labs > Van Aelst lab
Depositing User: CSHL Librarian
Date: August 2004
Date Deposited: 03 Feb 2012 15:35
Last Modified: 22 Feb 2017 21:07
PMCID: PMC446095
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