Interaction of Sesbania mosaic virus movement protein with the coat protein--implications for viral spread.
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.
Department of Biochemistry, Indian Institute of Science, Bangalore, India.
Soumya Roy
SR
Chowdhury
Handanahal Subbarao
HS
Savithri
D000595:Amino Acid Sequence; D036022:Capsid Proteins / Q000378:metabolism*; D002942:Circular Dichroism; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D008124:Locomotion / Q000502:physiology*; D008969:Molecular Sequence Data; D009029:Mosaic Viruses / Q000502:physiology*; D009598:Nitrophenylgalactosides / Q000378:metabolism; D053830:Plant Viral Movement Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D011485:Protein Binding / Q000187:drug effects / Q000502:physiology; D054730:Protein Interaction Domains and Motifs / Q000502:physiology; D055550:Protein Stability; D017433:Protein Structure, Secondary; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D011994:Recombinant Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D017384:Sequence Deletion / Q000502:physiology; D017386:Sequence Homology, Amino Acid; D049931:Sesbania / Q000821:virology*; D012965:Sodium Chloride / Q000494:pharmacology; D013050:Spectrometry, Fluorescence; D020798:Two-Hybrid System Techniques; D001616:beta-Galactosidase / Q000378:metabolism
D036022:Capsid Proteins; D053830:Plant Viral Movement Proteins; D011993:Recombinant Fusion Proteins; D011994:Recombinant Proteins; C055012:2-nitrophenylgalactoside; D009598:Nitrophenylgalactosides; D012965:Sodium Chloride; D001616:beta-Galactosidase