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چکیده
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The exploration of protein-protein interactions among viral proteins, as well as their interactions with host plant proteins, and their localization within infected plant cells, is crucial for inhibiting the formation of viral replication complexes and developing resistance in plant species against harmful viruses. The beet necrotic yellow vein virus (BNYVV), which induces rhizomania in sugar beets, is prevalent in sugar beet growing areas across Europe, Asia, and America. Field isolates of benyviruses typically consist of four RNA species, with RNA-5 being present in only certain BNYVV isolates, which appear to be more virulent. RNA-1 encodes a non-structural p237 polyprotein that is cleaved into two proteins (p150 and p66) by a cis-acting protease activity. The non-structural proteins of BNYVV share close homology with replication proteins of positive strand RNA viruses, such as Hepeviruses, rather than with replicases of other plant viruses. The p150 protein contains domains with methyltransferase, protease, helicase, and two domains of unknown function, while p66 encompasses the RNA-dependent RNA-polymerase signature. Through yeast two-hybrid analysis, colocalization with FRET-FLIM analyses, and co-immunoprecipitation, we have demonstrated an association of the p150 N-terminal domain with itself and with the internal domain of p66-RdRp, indicating their involvement in directing the Benyvirus replicase to the endoplasmic reticulum. These findings suggest the presence of a multimeric complex in the vicinity of the cellular membranous web. The N-terminal part of p237 is responsible for endoplasmic reticulum localization, as it possesses hydrophobic predicted transmembrane regions that could explain the anchoring of the replicase to the ER membrane.
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