title: Probing transcription factor diffusion dynamics in the living mammalian embryo with photoactivatable fluorescence correlation spectroscopy.
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.
authors.affiliation: European Molecular Biology Laboratory (EMBL) Australia, Australian Regenerative Medicine Institute, Monash University, Clayton, Victoria 3800, Australia.
authors.forename: Gurpreet
authors.initials: G
authors.lastname: Kaur
authors.forename: Mauro W
authors.initials: MW
authors.lastname: Costa
authors.forename: Christian M
authors.initials: CM
authors.lastname: Nefzger
authors.forename: Juan
authors.initials: J
authors.lastname: Silva
authors.forename: Juan Carlos
authors.initials: JC
authors.lastname: Fierro-González
authors.forename: Jose M
authors.initials: JM
authors.lastname: Polo
authors.forename: Toby D M
authors.initials: TD
authors.lastname: Bell
authors.forename: Nicolas
authors.initials: N
authors.lastname: Plachta
mesh_terms: D000818:Animals; D019070:Cell Lineage; D004058:Diffusion; D004622:Embryo, Mammalian / Q000166:cytology / Q000378:metabolism*; D005456:Fluorescent Dyes / Q000378:metabolism; D049452:Green Fluorescent Proteins / Q000378:metabolism; D051379:Mice; D013050:Spectrometry, Fluorescence / Q000379:methods*; D014157:Transcription Factors / Q000378:metabolism*
chemical_list: D005456:Fluorescent Dyes; D014157:Transcription Factors; D049452:Green Fluorescent Proteins
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