Probing transcription factor diffusion dynamics in the living mammalian embryo with photoactivatable fluorescence correlation spectroscopy

Gurpreet Kaur, Mauro W. Costa, Christian M. Nefzger, Juan Silva, Juan Carlos Fierro-González, Jose M. Polo, Toby D.M. Bell () and Nicolas Plachta ()
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Gurpreet Kaur: European Molecular Biology Laboratory (EMBL) Australia, Australian Regenerative Medicine Institute, Monash University
Mauro W. Costa: European Molecular Biology Laboratory (EMBL) Australia, Australian Regenerative Medicine Institute, Monash University
Christian M. Nefzger: Monash Immunology and Stem Cell Laboratories, Australian Regenerative Medicine Institute, Monash University
Juan Silva: European Molecular Biology Laboratory (EMBL) Australia, Australian Regenerative Medicine Institute, Monash University
Juan Carlos Fierro-González: European Molecular Biology Laboratory (EMBL) Australia, Australian Regenerative Medicine Institute, Monash University
Jose M. Polo: Monash Immunology and Stem Cell Laboratories, Australian Regenerative Medicine Institute, Monash University
Toby D.M. Bell: School of Chemistry, Monash University
Nicolas Plachta: European Molecular Biology Laboratory (EMBL) Australia, Australian Regenerative Medicine Institute, Monash University

Nature Communications, 2013, vol. 4, issue 1, 1-13

Abstract: Abstract Transcription factors use diffusion to search the DNA, yet the mechanisms controlling transcription factor diffusion during mammalian development remain poorly understood. Here we combine photoactivation and fluorescence correlation spectroscopy to study transcription factor diffusion in developing mouse embryos. We show that the pluripotency-associated transcription factor Oct4 displays both fast and Brownian and slower subdiffusive behaviours that are controlled by DNA interactions. Following cell lineage specification, the slower DNA-interacting diffusion fraction distinguishes pluripotent from extraembryonic cell nuclei. Similar to Oct4, Sox2 shows slower diffusion in pluripotent cells while Cdx2 displays opposite dynamics, suggesting that slow diffusion may represent a general feature of transcription factors in lineages where they are essential. Slow Oct4 subdiffusive behaviours are conserved in embryonic stem cells and induced pluripotent stem cells (iPS cells), and lost during differentiation. We also show that Oct4 diffusion depends on its interaction with ERG-associated protein with SET domain. Photoactivation and fluorescence correlation spectroscopy provides a new intravital approach to study transcription factor diffusion in complex in vivo systems.

Date: 2013
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DOI: 10.1038/ncomms2657

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