Abstract
Most plant viruses move between plant cells with the help of their movement proteins (MPs). MPs are multifunctional proteins, and one of their functions is almost invariably binding to nucleic acids. Presumably, the MP—nucleic acid interaction is directly involved in formation of nucleoprotein complexes that function as intermediates in the cell-to-cell transport of many plant viruses. Thus, when studying a viral MP, it is important to determine whether or not it binds nucleic acids, and to characterize the hallmark parameters of such binding, i.e., preference for single- or double-stranded nucleic acids and binding cooperativity and sequence specificity. Here, we present two major experimental approaches, native gel mobility shift assay and ultra violet (UV) light cross-linking, for detection and characterization of MP binding to DNA and RNA molecules. We also describe protocols for purification of recombinant viral MPs over-expressed in bacteria and production of different DNA and RNA probes for these binding assays.
Keywords
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Acknowledgments
We apologize to colleagues whose original works have not been cited due to the lack of space. The work in our laboratories is supported by grants from NIH, NSF, USDA, BARD, and BSF to VC, and by grants from BARD and HFSP to TT.
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Tzfira, T., Citovsky, V. (2008). Probing Interactions Between Plant Virus Movement Proteins and Nucleic Acids. In: Foster, G.D., Johansen, I.E., Hong, Y., Nagy, P.D. (eds) Plant Virology Protocols. Methods in Molecular Biology™, vol 451. Humana Press. https://doi.org/10.1007/978-1-59745-102-4_21
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DOI: https://doi.org/10.1007/978-1-59745-102-4_21
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