Interaction of Sesbania mosaic virus movement protein with the coat protein - implications for viral spread

• Published in: The FEBS journal 2011, Vol.278 (2), p.257-272
• Main Authors: Chowdhury, Soumya Roy, Savithri, Handanahal Subbarao
• Format: Article
• Language: English
• Subjects: Amino Acid Sequence; amino acids; beta-Galactosidase - metabolism; Biochemistry; Capsid Proteins - metabolism; cells; Circular Dichroism; coat protein; coat proteins; enzyme-linked immunosorbent assay; Escherichia coli; Escherichia coli - genetics; Escherichia coli - metabolism; Gene expression; gene overexpression; genes; Hydrogen-Ion Concentration; Locomotion - physiology; Molecular Sequence Data; Mosaic Viruses - physiology; mutants; Mutation; Nitrophenylgalactosides - metabolism; Plant Viral Movement Proteins - chemistry; Plant Viral Movement Proteins - genetics; Plant Viral Movement Proteins - metabolism; Protein Binding - drug effects; Protein Binding - physiology; Protein Interaction Domains and Motifs - physiology; Protein Stability; Protein Structure, Secondary; Proteins; protein–protein interaction; Recombinant Fusion Proteins - genetics; Recombinant Fusion Proteins - isolation & purification; Recombinant Fusion Proteins - metabolism; recombinant movement protein; Recombinant Proteins - genetics; Recombinant Proteins - isolation & purification; Recombinant Proteins - metabolism; RNA; Sequence Deletion - physiology; sequence homology; Sequence Homology, Amino Acid; Sesbania; Sesbania - virology; Sesbania mosaic virus; sobemovirus; Sobemoviruses; Sodium Chloride - pharmacology; Southern bean mosaic virus; Spectrometry, Fluorescence; Two-Hybrid System Techniques; Viruses
• ISSN: 1742-464X; 1742-4658
• DOI: 10.1111/j.1742-4658.2010.07943.x

Sesbania mosaic virus (SeMV) is a single-stranded positive-sense RNA plant virus belonging to the genus Sobemovirus. The movement protein (MP) encoded by SeMV ORF1 showed no significant sequence similarity with MPs of other genera, but showed 32% identity with the MP of Southern bean mosaic virus within the Sobemovirus genus. With a view to understanding the mechanism of cell-to-cell movement in sobemoviruses, the SeMV MP gene was cloned, over-expressed in Escherichia coli and purified. Interaction of the recombinant MP with the native virus (NV) was investigated by ELISA and pull-down assays. It was observed that SeMV MP interacted with NV in a concentration- and pH-dependent manner. Analysis of N- and C-terminal deletion mutants of the MP showed that SeMV MP interacts with the NV through the N-terminal 49 amino acid segment. Yeast two-hybrid assays confirmed the in vitro observations, and suggested that SeMV might belong to the class of viruses that require MP and NV/coat protein for cell-to-cell movement.