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

Natalie M. Clark; Elizabeth Hinde; Cara M. Winter; Adam P. Fisher; Giuseppe Crosti; Ikram Blilou; Enrico Gratton; Philip N. Benfey; Rosangela Sozzani

doi:10.7554/eLife.14770

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

Natalie M. Clark, Elizabeth Hinde, Cara M. Winter, Adam P. Fisher, Giuseppe Crosti, Ikram Blilou, Enrico Gratton, Philip N. Benfey, Rosangela Sozzani^*

^*Corresponding author for this work

Research output: Contribution to journal › Article › peer-review

64 Scopus citations

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.

Original language	English (US)
Article number	e14770
Journal	eLife
Volume	5
Issue number	JUN2016
DOIs	https://doi.org/10.7554/eLife.14770
State	Published - Jun 11 2016
Externally published	Yes

Bibliographical note

Funding Information:
National Science Foundation MCB-1453130 Rosangela Sozzani. National Science Foundation DGE-1252376 Natalie M Clark, Adam P Fisher. National Institutes of Health GM-106690-01 Cara M Winter. National Institutes of Health P41-GM103540 Enrico Gratton. National Institutes of Health P50-GM076516 Enrico Gratton. National Science Foundation IOS-1021619 Philip N Benfey. National Institutes of Health R01-GM043778 Philip N Benfey. Howard Hughes Medical Institute GBMF3405 Philip N Benfey. Gordon and Betty Moore Foundation GBMF3405 Philip N Benfey. The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.

Publisher Copyright:
© Clark et al.

ASJC Scopus subject areas

Neuroscience(all)
Immunology and Microbiology(all)
Biochemistry, Genetics and Molecular Biology(all)

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10.7554/eLife.14770

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title = "Tracking transcription factor mobility and interaction in arabidopsis roots with fluorescence correlation spectroscopy",

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.",

author = "Clark, {Natalie M.} and Elizabeth Hinde and Winter, {Cara M.} and Fisher, {Adam P.} and Giuseppe Crosti and Ikram Blilou and Enrico Gratton and Benfey, {Philip N.} and Rosangela Sozzani",

note = "Funding Information: National Science Foundation MCB-1453130 Rosangela Sozzani. National Science Foundation DGE-1252376 Natalie M Clark, Adam P Fisher. National Institutes of Health GM-106690-01 Cara M Winter. National Institutes of Health P41-GM103540 Enrico Gratton. National Institutes of Health P50-GM076516 Enrico Gratton. National Science Foundation IOS-1021619 Philip N Benfey. National Institutes of Health R01-GM043778 Philip N Benfey. Howard Hughes Medical Institute GBMF3405 Philip N Benfey. Gordon and Betty Moore Foundation GBMF3405 Philip N Benfey. The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication. Publisher Copyright: {\textcopyright} Clark et al.",

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

Abstract

Bibliographical note

ASJC Scopus subject areas

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