Structural characterization of stem cell factors Oct4, Sox2, Nanog and Esrrb disordered domains, and a method to identify their phospho-dependent binding partners

Abstract

The combined expression of a handful of pluripotency transcription factors (PluriTFs) in 24 somatic cells can generate induced pluripotent stem cells (iPSCs). Here, we report the 25 structural characterization of disordered regions contained in four important PluriTFs, namely 26 Oct4, Sox2, Nanog and Esrrb. Moreover, many post-translational modifications (PTMs) have 27 been detected on PluriTFs, whose roles are not yet characterized. To help in their study, we 28 also present a method i) to produce well-characterized phosphorylation states of PluriTFs, 29 using NMR analysis, and ii) to use them for pull-downs in stem cell extracts analyzed by 30 quantitative proteomics to identify of Sox2 binders.