1. Developmental Biology
  2. Plant Biology

Tracking transcription factor mobility and interaction in Arabidopsis roots with fluorescence correlation spectroscopy

  1. Natalie M Clark
  2. Elizabeth Hinde
  3. Cara M Winter
  4. Adam P Fisher
  5. Giuseppe Crosti
  6. Ikram Blilou
  7. Enrico Gratton
  8. Philip N Benfey  Is a corresponding author
  9. Rosangela Sozzani
  1. North Carolina State University, United States
  2. University of California, Irvine, United States
  3. Howard Hughes Medical Institute, Duke University, United States
  4. Wageningen University, Netherlands
https://doi.org/10.7554/eLife.14770
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Cite this article
  1. Natalie M Clark
  2. Elizabeth Hinde
  3. Cara M Winter
  4. Adam P Fisher
  5. Giuseppe Crosti
  6. Ikram Blilou
  7. Enrico Gratton
  8. Philip N Benfey
  9. Rosangela Sozzani
(2016)
Tracking transcription factor mobility and interaction in Arabidopsis roots with fluorescence correlation spectroscopy
eLife 5:e14770.
https://doi.org/10.7554/eLife.14770
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  3. Download .RIS

Abstract

To understand complex regulatory processes in multicellular organisms, it is critical to be able to quantitatively analyze protein movement and protein-protein interactions in time and space. During Arabidopsis development, the intercellular movement of SHORTROOT (SHR) and subsequent interaction with its downstream target SCARECROW (SCR) control root patterning and cell fate specification. However, quantitative information about the spatio-temporal dynamics of SHR movement and SHR-SCR interaction is currently unavailable. Here, we quantify parameters including SHR mobility, oligomeric state, and association with SCR using a combination of Fluorescent Correlation Spectroscopy (FCS) techniques. We then incorporate these parameters into a mathematical model of SHR and SCR, which shows that SHR reaches a steady state in minutes, while SCR and the SHR-SCR complex reach a steady-state between 18 and 24 hours. Our model reveals the timing of SHR and SCR dynamics and allows us to understand how protein movement and protein-protein stoichiometry contribute to development.

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Author details

  1. Natalie M Clark

    Department of Plant and Microbial Biology, North Carolina State University, Raleigh, United States
    Competing interests
    The authors declare that no competing interests exist.

  2. Elizabeth Hinde

    Laboratory for Fluorescence Dynamics, University of California, Irvine, Irvine, United States
    Competing interests
    The authors declare that no competing interests exist.

  3. Cara M Winter

    Department of Biology, Howard Hughes Medical Institute, Duke University, Durham, United States
    Competing interests
    The authors declare that no competing interests exist.

  4. Adam P Fisher

    Department of Plant and Microbial Biology, North Carolina State University, Raleigh, United States
    Competing interests
    The authors declare that no competing interests exist.

  5. Giuseppe Crosti

    Department of Biology, Howard Hughes Medical Institute, Duke University, Durham, United States
    Competing interests
    The authors declare that no competing interests exist.

  6. Ikram Blilou

    Plant Developmental Biology, Wageningen University, Wageningen, Netherlands
    Competing interests
    The authors declare that no competing interests exist.

  7. Enrico Gratton

    Laboratory for Fluorescence Dynamics, University of California, Irvine, Irvine, United States
    Competing interests
    The authors declare that no competing interests exist.

  8. Philip N Benfey

    Department of Biology, Howard Hughes Medical Institute, Duke University, Durham, United States
    For correspondence
    philip.benfey@duke.edu
    Competing interests
    The authors declare that no competing interests exist.

  9. Rosangela Sozzani

    Department of Plant and Microbial Biology, North Carolina State University, Raleigh, United States
    Competing interests
    The authors declare that no competing interests exist.

Copyright

© 2016, Clark et al.

This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.

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  1. Natalie M Clark
  2. Elizabeth Hinde
  3. Cara M Winter
  4. Adam P Fisher
  5. Giuseppe Crosti
  6. Ikram Blilou
  7. Enrico Gratton
  8. Philip N Benfey
  9. Rosangela Sozzani
(2016)
Tracking transcription factor mobility and interaction in Arabidopsis roots with fluorescence correlation spectroscopy
eLife 5:e14770.
https://doi.org/10.7554/eLife.14770

Share this article

https://doi.org/10.7554/eLife.14770

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