title: The RNA-dependent RNA polymerase of Citrus tristeza virus forms oligomers.
abstract: The RNA-dependent RNA polymerases (RdRp) from Citrus tristeza virus (CTV) were tagged with HA and FLAG epitopes. Differentially tagged proteins were expressed either individually or concomitantly in Escherichia coli. Immunoprecipitation of the expressed proteins with anti-FLAG antibody followed by Western blot with anti-HA antibody demonstrated that molecules of RdRp from CTV interact to form oligomers. Yeast two-hybrid assays showed that molecules of RdRp interact in eukaryotic cells. Co-immunoprecipitation with anti-FLAG antibody of truncated HA-tagged RdRps (RdRpΔ1-166-HA, RdRpΔ1-390-HA, RdRp1-169-HA) co-expressed with full-length RdRp-FLAG showed that only RdRp1-169-HA interacted with the full-length FLAG-RdRp. Yeast two-hybrid assays with truncated RdRp constructs confirmed that the oligomerization site resides in the N-terminal region and that the first 169 aa of CTV RdRp are necessary and sufficient for oligomerization both in bacterial and yeast cells. Development of control strategies targeting viral RdRp oligomer formation may inhibit virus replication and prove useful in control of CTV.
authors.affiliation: Suleyman Demirel University, Faculty of Agriculture, Department of Plant Protection, Isparta 32260, Turkey. Electronic address: bayramcevik@sdu.edu.tr.
authors.forename: Bayram
authors.initials: B
authors.lastname: Cevik
mesh_terms: D015153:Blotting, Western; D002957:Citrus / Q000821:virology; D017790:Closterovirus / Q000201:enzymology*; D004926:Escherichia coli / Q000235:genetics; D047468:Immunoprecipitation; D055503:Protein Multimerization; D012324:RNA-Dependent RNA Polymerase / Q000378:metabolism*; D012441:Saccharomyces cerevisiae / Q000235:genetics; D020798:Two-Hybrid System Techniques
chemical_list: D012324:RNA-Dependent RNA Polymerase
keywords: CTV; Coimmunoprecipitation; Epitope tagging; Immunoblotting; Oligomerization; Protein expression; Protein–protein interaction; RdRp; Yeast-two-hybrid system