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

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

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Plant Developmental Biology, Wageningen University and Research Centre, Droevendaalsesteeg 1, Wageningen 6708PB, The Netherlands.
Molecular Genetics, Department Biology, Utrecht University, Padualaan 8, Utrecht 3581CH, The Netherlands.
CEPLAS (Cluster of Excellence on Plant Sciences), and Institute for Developmental Genetics, Heinrich Heine University, Universitätsstraße 1, Düsseldorf 40225, Germany.
Center for Advanced Imaging, Heinrich Heine University, Universitätsstraße 1, Düsseldorf 40225, Germany.
Swammerdam Institute for Life Sciences, Section of Molecular Cytology, van Leeuwenhoek Centre for Advanced Microscopy, University of Amsterdam, Science Park 904, Amsterdam 1098 XH, The Netherlands.
Departamento de Bioquímica, UAM, Instituto de Investigaciones Biomédicas Alberto Sols, CSIC-UAM, Arturo Duperier 4, 28029 Madrid, Spain.


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.

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