Gel mobility shift assays for rna binding viral rnai suppressors

Tibor Csorba, J. Burgyán

Research output: Chapter in Book/Report/Conference proceedingChapter

3 Citations (Scopus)

Abstract

The host-virus interaction is a continuous coevolutionary race involving both host defence strategies and virus escape mechanisms. RNA silencing is one of the main processes employed by eukaryotic organisms to fight viruses. However, viruses encode suppressor proteins to counteract this antiviral mechanism. Virtually all plant viruses encode at least one suppressor. In spite of being highly diverse at the protein level, a large group of these proteins inhibit RNA silencing very similarly, by sequestration of double-stranded RNA or small-interfering RNA molecules, the central players of the pathway. The RNA binding capacity of virus suppressor proteins can be studied by the electrophoretic mobility shift assay method. Also known as gel retardation assay, gel mobility assay, gel shift assay or band shift assay, EMSA is an in vitro technique used to characterize protein:DNA or protein:RNA interactions. The method had been developed based on the observation that protein: nucleic acid complexes migrate slower through a non-denaturing polyacrylamide gel than the free nucleic acid fragments. Here, we provide a detailed protocol for the analysis of crucifer-infecting Tobacco mosaic tobamovirus (cr-TMV) silencing suppressor protein p122 RNA binding capacity.

Original languageEnglish
Title of host publicationMethods in Molecular Biology
Pages245-252
Number of pages8
Volume721
DOIs
Publication statusPublished - 2011

Publication series

NameMethods in Molecular Biology
Volume721
ISSN (Print)10643745

Fingerprint

Viral RNA
Electrophoretic Mobility Shift Assay
Gels
Proteins
Viruses
RNA Interference
Nucleic Acids
Tobamovirus
RNA
Virus Attachment
Plant Viruses
RNA-Binding Proteins
Double-Stranded RNA
Small Interfering RNA
Tobacco
Antiviral Agents
DNA

Keywords

  • Antiviral silencing
  • Band shift assay
  • cr-TMV p122
  • Electro mobility shift assay
  • RNA binding
  • siRNA
  • Viral suppressor proteins

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics
  • Medicine(all)

Cite this

Csorba, T., & Burgyán, J. (2011). Gel mobility shift assays for rna binding viral rnai suppressors. In Methods in Molecular Biology (Vol. 721, pp. 245-252). (Methods in Molecular Biology; Vol. 721). https://doi.org/10.1007/978-1-61779-037-9_15

Gel mobility shift assays for rna binding viral rnai suppressors. / Csorba, Tibor; Burgyán, J.

Methods in Molecular Biology. Vol. 721 2011. p. 245-252 (Methods in Molecular Biology; Vol. 721).

Research output: Chapter in Book/Report/Conference proceedingChapter

Csorba, T & Burgyán, J 2011, Gel mobility shift assays for rna binding viral rnai suppressors. in Methods in Molecular Biology. vol. 721, Methods in Molecular Biology, vol. 721, pp. 245-252. https://doi.org/10.1007/978-1-61779-037-9_15
Csorba T, Burgyán J. Gel mobility shift assays for rna binding viral rnai suppressors. In Methods in Molecular Biology. Vol. 721. 2011. p. 245-252. (Methods in Molecular Biology). https://doi.org/10.1007/978-1-61779-037-9_15
Csorba, Tibor ; Burgyán, J. / Gel mobility shift assays for rna binding viral rnai suppressors. Methods in Molecular Biology. Vol. 721 2011. pp. 245-252 (Methods in Molecular Biology).
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