Fluorescent protein recruitment assay for demonstration and analysis of in vivo protein interactions in plant cells and its application to Tobacco mosaic virus movement protein

Plant J. 2010 Apr 1;62(1):171-7. doi: 10.1111/j.1365-313X.2010.04126.x. Epub 2010 Jan 7.

Abstract

We describe a simple fluorescent protein-based method to investigate interactions with a viral movement protein in living cells that relies on the in vivo re-localization of proteins in the presence of their interaction partners. We apply this method in combination with fluorescence lifetime imaging microscopy (FLIM) to demonstrate that a domain of the Tobacco mosaic virus (TMV) movement protein (MP) previously predicted to mediate protein:protein interactions is dispensable for these contacts. We suggest that this method can be generalized for analysis of other protein interactions in planta.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Luminescent Proteins / metabolism
  • Microscopy, Confocal
  • Microscopy, Fluorescence*
  • Nicotiana / genetics
  • Nicotiana / virology*
  • Plant Viral Movement Proteins / metabolism*
  • Plants, Genetically Modified / genetics
  • Plants, Genetically Modified / virology
  • Protein Interaction Mapping*
  • Tobacco Mosaic Virus / metabolism*

Substances

  • Luminescent Proteins
  • Plant Viral Movement Proteins