In vivo FRET-FLIM reveals cell-type-specific protein interactions in Arabidopsis roots

Nature. 2017 Aug 3;548(7665):97-102. doi: 10.1038/nature23317. Epub 2017 Jul 26.


During multicellular development, specification of distinct cell fates is often regulated by the same transcription factors operating differently in distinct cis-regulatory modules, either through different protein complexes, conformational modification of protein complexes, or combinations of both. Direct visualization of different transcription factor complex states guiding specific gene expression programs has been challenging. Here we use in vivo FRET-FLIM (Förster resonance energy transfer measured by fluorescence lifetime microscopy) to reveal spatial partitioning of protein interactions in relation to specification of cell fate. We show that, in Arabidopsis roots, three fully functional fluorescently tagged cell fate regulators establish cell-type-specific interactions at endogenous expression levels and can form higher order complexes. We reveal that cell-type-specific in vivo FRET-FLIM distributions reflect conformational changes of these complexes to differentially regulate target genes and specify distinct cell fates.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Arabidopsis / cytology*
  • Arabidopsis / metabolism*
  • Arabidopsis Proteins / genetics
  • Arabidopsis Proteins / metabolism
  • Carrier Proteins / genetics
  • Carrier Proteins / metabolism
  • Cell Lineage
  • Endoderm / cytology
  • Endoderm / metabolism
  • Fluorescence Resonance Energy Transfer*
  • HeLa Cells
  • Homeodomain Proteins / genetics
  • Humans
  • Microscopy, Fluorescence
  • Mutation
  • Organ Specificity
  • Plant Roots / cytology*
  • Plant Roots / metabolism*
  • Protein Binding
  • Protein Interaction Mapping / methods*
  • Protein Interaction Maps*
  • Stem Cells / cytology
  • Stem Cells / metabolism
  • Transcription Factors / metabolism


  • Arabidopsis Proteins
  • Carrier Proteins
  • Homeodomain Proteins
  • JACKDAW protein, Arabidopsis
  • NUC protein, Arabidopsis
  • SCR protein, Arabidopsis
  • SHORT ROOT protein, Arabidopsis
  • Transcription Factors
  • WUSCHEL protein, Arabidopsis