Molecular anatomy of plant viral genome replication machinery for the development of cell-free viral replication system.

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This study was intended to analyze the molecular structure of viral replication complexes (VRC) of 5 different plant viruses for developing the cell-free viral replication systems.1.Molecular anatomy of TMV replication complex :Tobacco mosaic virus RNA dependent RNA polymerase (RdRp) was shown to interact specifically with the host protein kinase (NtPERK1), protein phosphatase 2C (NtPP2C-2), arginine decarboxylase (ADC) and ring-finger protein (PHF15) those might regulate the viral replication. Also elongation factor eEF1A was shown to bind directly with TMV RdRp as well as 3'-UTR of TMV RNA, which indicated that eEF1A is one of the essential factors of VRC.2.Molecular anatomy of SbCMV replication complex :ORF II product of Soybean chlorotic mottle virus (SbCMV) showed the self-interaction that was indispensable to viral pathogenicity.3.Molecular anatomy of ASGV replication complex :Apple stem grooving virus (ASGV) was revealed to express its proteins through the frameshift strategy and the variable region was shown to be involved in its pathogenicity.4.Molecular anatomy of TVX replication complex :Potato virus X belonging to the same genus of Tulip virus X (TVX) showed that the C-terminus region of the replicase and 5'untranslated region were responsible for different types of symptoms.5.Molecular anatomy of SBWMV replication complex and trial for developing cell-free replication system :Replication of Soil-borne wheat mosaic virus (SBWMV) was more efficient at 17℃ than at 15℃ or 20℃. The p19 cysteine-rich protein, an RNA silencing suppressor, facilitated RNA accumulation in transfected cells late in infection as well as cell-to-cell movement of viral RNA. SBWMV RNA1 was translated in vitro in transcription-coupled wheat germ extracts using RNA1 cDNA with the upstream T7 promoter as a template. It was confirmed that translational readthrough at the UGA termination codon of the p152 gene could occur as in vivo at 24℃.