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 journalArticlepeer-review

45 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 languageEnglish (US)
Article numbere14770
JournaleLife
Volume5
Issue numberJUN2016
DOIs
StatePublished - Jun 11 2016

ASJC Scopus subject areas

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

Fingerprint Dive into the research topics of 'Tracking transcription factor mobility and interaction in arabidopsis roots with fluorescence correlation spectroscopy'. Together they form a unique fingerprint.

Cite this