TWIST1 and chromatin regulatory proteins interact to guide neural crest cell differentiation
- CMRI, The University of Sydney, Australia
Abstract
Protein interaction is critical molecular regulatory activity underlining cellular functions and precise cell fate choices. Using TWIST1 BioID-proximity-labelling and network propagation analyses, we discovered and characterized a TWIST-chromatin regulatory module (TWIST1-CRM) in the neural crest cells (NCC). Combinatorial perturbation of core members of TWIST1-CRM: TWIST1, CHD7, CHD8, and WHSC1 in cell models and mouse embryos revealed that loss of the function of the regulatory module resulted in abnormal differentiation of NCCs and compromised craniofacial tissue patterning. Following NCC delamination, low level of TWIST1-CRM activity is instrumental to stabilize the early NCC signatures and migratory potential by repressing the neural stem cell programs. High level of TWIST1 module activity at later phases commits the cells to the ectomesenchyme. Our study further revealed the functional interdependency of TWIST1 and potential neurocristopathy factors in NCC development.
Data availability
All data generated or analyzed during this study are included in the manuscript and supporting files. Sequencing data have been deposited in GEO under accession codes GSE130251. External data analyzed has been listed in Supplementary File 6.
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TWIST1 direct targets during embryonic stem cell differentiation [ChIP-seq]NCBI Gene Expression Omnibus, GSE130251.
-
Dissecting neural differentiation regulatory networks through epigenetic footprintingNCBI Gene Expression Omnibus, GSE62193.
-
AF22_H3K36me3NCBI Gene Expression Omnibus, GSM2902410.
-
Genome-wide maps of chromatin state during the differentiation of hESC into hNECs (ChIP-Seq)NCBI Gene Expression Omnibus, GSM1973975.
-
Transcription factors interfering with dedifferentiation induce direct conversionNCBI Gene Expression Omnibus, GSM1012189.
-
Selective influence of Sox2 on POU transcription factor binding in embryonic and neural stem cellsNCBI Gene Expression Omnibus, GSM1711445.
-
Neural Progenitors Adopt Specific Identities by Directly Repressing All Alternative Progenitor Transcriptional ProgramsThe European Nucleotide Archive, ERS580651.
-
Spatio-temporal structure of cell fate decisions in murine neural crestNCBI Gene Expression Omnibus, GSE129114.
Article and author information
Author details
Funding
National Health and Medical Research Council (1066832,1079160,1003100,1110751)
- Mark E Graham
- Patrick PL Tam
Australian Research Council (1094008)
- Xiaochen Fan
- Patrick PL Tam
University of Sydney
- Xiaochen Fan
Children's Medical Research Institute
- Xiaochen Fan
- Pierre Osteil
- Nicolas Fossat
Carlsbergfondet (CF15-1056,CF16-0066)
- Kasper Engholm-Keller
The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.
Ethics
Animal experimentation: Animal experimentations were performed in compliance with animal ethics and welfare guidelines stipulated by the Children's Medical Research Institute/Children's Hospital at Westmead Animal Ethics Committee, protocol number C230.
Copyright
© 2021, Fan et al.
This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.
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TWIST1 and chromatin regulatory proteins interact to guide neural crest cell differentiation
eLife 10:e62873.
https://doi.org/10.7554/eLife.62873
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