The CTD of EBF1 enhances Brg1 recruitment to chromatin
Ontology highlight
ABSTRACT: EBF1 binds lymphoid progenitor chromatin prior to the detection of chromatin accessibility, whereby a C-terminal domain (CTD) enables “pioneering” independently of cooperating transcription factors. The CTD of EBF1 includes a short prion-like domain (PLD) with an ability of liquid-liquid phase separation in vitro and in vivo. The PLD of EBF1 mediates the recruitment of the chromatin remodeler Brg1 (a subunit of SWI/SNF remodeling complex).
Project description:EBF1 is a transcription factor crucial for early B cell development. The phase separation ability of the EBF1 C-terminal domain (CTD) is essential for the transcription activation and pioneering function of EBF1. We identified the specific tyrosine residues in the CTD required for phase separation. We transduced pro-B cells with EBF1 wt, deletion of the entire CTD (dC), or mutation of these tyrosines (YmA). We performed ChIP-seq, ATAC-seq, and RNA-seq to see the effect of disrupting phase separation ability on DNA binding, chromatin opening, and target gene expression. Our findings confirm that phase separation is important for the EBF1 function.
Project description:EBF1 is a transcription factor crucial for early B cell development. The phase separation ability of the EBF1 C-terminal domain (CTD) is essential for the transcription activation and pioneering function of EBF1. We identified the specific tyrosine residues in the CTD required for phase separation. We transduced pro-B cells with EBF1 wt, deletion of the entire CTD (dC), or mutation of these tyrosines (YmA). We performed ChIP-seq, ATAC-seq, and RNA-seq to see the effect of disrupting phase separation ability on DNA binding, chromatin opening, and target gene expression. Our findings confirm that phase separation is important for the EBF1 function.
Project description:EBF1 is a transcription factor crucial for early B cell development. The phase separation ability of the EBF1 C-terminal domain (CTD) is essential for the transcription activation and pioneering function of EBF1. We identified the specific tyrosine residues in the CTD required for phase separation. We transduced pro-B cells with EBF1 wt, deletion of the entire CTD (dC), or mutation of these tyrosines (YmA). We performed ChIP-seq, ATAC-seq, and RNA-seq to see the effect of disrupting phase separation ability on DNA binding, chromatin opening, and target gene expression. Our findings confirm that phase separation is important for the EBF1 function.
Project description:EBF1 is essential for B cell specification and commitment. To explore the dynamics of EBF1 initiated B cell programming, we performed EBF1 ChIP-seq, ATAC-seq, bisulfite-seq, RNA-seq and several histone ChIP-seq analyses at different stages of the transition from Ebf1-/- pre-pro-B to pro-B triggered by EBF1 restoration. We also performed Pax5 ChIP-seq in Ebf1-/- pre-pro-B cell and EBF1-restored pro-B cell to study the pioneering function of EBF1 that allows other transcription factors to access certain chromatin sites.
Project description:EBF1 is essential for B cell specification and commitment. To explore the dynamics of EBF1 initiated B cell programming, we performed EBF1 ChIP-seq, ATAC-seq, bisulfite-seq, RNA-seq and several histone ChIP-seq analyses at different stages of the transition from Ebf1-/- pre-pro-B to pro-B triggered by EBF1 restoration. We also performed Pax5 ChIP-seq in Ebf1-/- pre-pro-B cell and EBF1-restored pro-B cell to study the pioneering function of EBF1 that allows other transcription factors to access certain chromatin sites.
Project description:EBF1 is essential for B cell specification and commitment. To explore the dynamics of EBF1 initiated B cell programming, we performed EBF1 ChIP-seq, ATAC-seq, bisulfite-seq, RNA-seq and several histone ChIP-seq analyses at different stages of the transition from Ebf1-/- pre-pro-B to pro-B triggered by EBF1 restoration. We also performed Pax5 ChIP-seq in Ebf1-/- pre-pro-B cell and EBF1-restored pro-B cell to study the pioneering function of EBF1 that allows other transcription factors to access certain chromatin sites.
Project description:EBF1 is essential for B cell specification and commitment. To explore the dynamics of EBF1 initiated B cell programming, we performed EBF1 ChIP-seq, ATAC-seq, bisulfite-seq, RNA-seq and several histone ChIP-seq analyses at different stages of the transition from Ebf1-/- pre-pro-B to pro-B triggered by EBF1 restoration. We also performed Pax5 ChIP-seq in Ebf1-/- pre-pro-B cell and EBF1-restored pro-B cell to study the pioneering function of EBF1 that allows other transcription factors to access certain chromatin sites.
Project description:EBF1 is essential for B cell specification and commitment. To explore the dynamics of EBF1 initiated B cell programming, we performed EBF1 ChIP-seq, ATAC-seq, bisulfite-seq, RNA-seq and several histone ChIP-seq analyses at different stages of the transition from Ebf1-/- pre-pro-B to pro-B triggered by EBF1 restoration. We also performed Pax5 ChIP-seq in Ebf1-/- pre-pro-B cell and EBF1-restored pro-B cell to study the pioneering function of EBF1 that allows other transcription factors to access certain chromatin sites.
Project description:EBF1 is essential for B cell specification and commitment. To explore the dynamics of EBF1 initiated B cell programming, we performed EBF1 ChIP-seq, ATAC-seq, bisulfite-seq, RNA-seq and several histone ChIP-seq analyses at different stages of the transition from Ebf1-/- pre-pro-B to pro-B triggered by EBF1 restoration. We also performed Pax5 ChIP-seq in Ebf1-/- pre-pro-B cell and EBF1-restored pro-B cell to study the pioneering function of EBF1 that allows other transcription factors to access certain chromatin sites.