An extraction method for tobacco mosaic virus movement protein localizing in plasmodesmata

Protoplasma. 2005 Apr;225(1-2):85-92. doi: 10.1007/s00709-004-0080-4. Epub 2005 May 4.

Abstract

The intercellular communication by plasmodesmata (PD) is important for the growth and development of plants, and the transport of macromolecules through PD is likely to be regulated by developmental signals. While PD in the apical meristem transport macromolecules such as mRNAs, the branched PD in the mature leaf do not transport large macromolecules freely. The changes in PD during development might be important for sink-to-source changes in leaves, but the molecular mechanism is still unknown. Movement proteins (MPs) of the tobacco mosaic virus localize in the branched PD and increase the size exclusion limit, allowing transport of viral RNA. We developed a method for differential extraction of MP from isolated cell walls of transgenic tobacco leaves expressing MP or MP tagged with green-fluorescent protein. Lithium chloride at a concentration of 8 M removed filamentous structures in branched PD, the possible attachment site of MP. As some endogenous proteins were coeluted with MP by the treatment, this extraction method might be a powerful tool for investigating MP-interacting proteins in branched PD.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Green Fluorescent Proteins / genetics
  • Green Fluorescent Proteins / isolation & purification
  • Lithium Chloride
  • Microscopy, Electron
  • Plant Viral Movement Proteins
  • Plants, Genetically Modified
  • Plasmodesmata / chemistry
  • Recombinant Fusion Proteins / genetics
  • Recombinant Fusion Proteins / isolation & purification
  • Tobacco / chemistry
  • Tobacco / genetics
  • Tobacco / ultrastructure
  • Tobacco / virology
  • Tobacco Mosaic Virus / chemistry*
  • Tobacco Mosaic Virus / genetics
  • Viral Proteins / genetics
  • Viral Proteins / isolation & purification*

Substances

  • Plant Viral Movement Proteins
  • Recombinant Fusion Proteins
  • Viral Proteins
  • Green Fluorescent Proteins
  • Lithium Chloride