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

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    Author
    Clark, NM; Hinde, E; Winter, CM; Fisher, AP; Crosti, G; Blilou, I; Gratton, E; Benfey, PN; Sozzani, R
    Date
    2016-06-11
    Source Title
    eLife
    Publisher
    eLife Sciences Publications, Ltd
    University of Melbourne Author/s
    Hinde, Elizabeth
    Affiliation
    School of Physics
    Metadata
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    Document Type
    Journal Article
    Citations
    Clark, N. M., Hinde, E., Winter, C. M., Fisher, A. P., Crosti, G., Blilou, I., Gratton, E., Benfey, P. N. & Sozzani, R. (2016). Tracking transcription factor mobility and interaction in Arabidopsis roots with fluorescence correlation spectroscopy.. Elife, 5 (JUN2016), pp.e14770-. https://doi.org/10.7554/eLife.14770.
    Access Status
    Open Access
    URI
    http://hdl.handle.net/11343/255085
    DOI
    10.7554/eLife.14770
    Open Access at PMC
    http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4946880
    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 hr. 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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