Analysis of pluripotency transcription factor interactions reveals preferential binding of NANOG to SOX2 rather than NANOG or OCT4

Abstract

The pluripotency transcription factors (TFs) Nanog, Sox2, and Oct4 are at the centre of the gene regulatory network that controls cell identity in embryonic stem (ES) cells. However, the mechanisms by which these factors control cell fate, and their interactions with one another are not fully understood. Here we combine biophysical and novel biochemical assays to assess how these factors interact with each other quantitatively. A new confocal microscopy method to detect binding of a target protein to a fluorescently labelled partner (coimmunoprecipitation bead imaging microscopy [CBIM]) is presented and used to demonstrate homotypic binding of Nanog and heterotypic binding between Nanog and Sox2 and between Nanog and Oct4. Using fluorescence correlation spectroscopy we show that in solution, Nanog but not Oct4 or Sox2 can form homodimers. Fluorescence Cross Correlation Spectroscopy shows that the affinity of Nanog for dimer formation is in the order Sox2 > Nanog > Oct4. Importantly, live cell analysis demonstrate

Mecanismos Moleculares Involucrados en La Diferenciación De Células Madre en Linajes Gliales

“…Lo que sí es importante dejar en claro, es que a nivel molecular las ESC expresan un conjunto de genes característicos del estado pluripotente, incluidos factores de transcripción como Oct-3/4, Sox2 y Nanog que, al ser expresados, permiten la indiferenciación , la autorrenovación y la potencia celular (Kashyap et al, 2009;Mistri et al, 2020;Sedaghat et al, 2019;Tremble et al, 2020) . La regulación de estos Factores de Transcripción (TF), sumado a los factores de regulación, determinan la diferenciación de linajes celulares, al inhibir o potenciar la expresión en las células pluripotentes embrionarias Ferreira et al, 2018;Taupin, 2010;Wang et al, 2006).…”

“…Las células madre pluripotentes (PSC), incluidas las ESC y las células madre pluripotentes inducidas (iPSC), se caracterizan por su capacidad para retener un amplio potencial de diferenciación, luego de períodos prolongados de cultivo. Esta propiedad de autorrenovación se mantiene mediante controles del ciclo celular que mantienen la capacidad proliferativa a largo plazo (Mistri et al, 2020;Sharma & Bhonde, 2020).…”