CREST functions as a molecular bridge between neuron-specific enhancers and epigenetic regulators in mature motor neurons

Motor neuron development requires precise transcriptional regulation mediated in part by BAF (BRG1/BRM-associated factor) chromatin remodeling complexes. The neuron-specific BAF subunit CREST is critical for neuronal function and is mutated in some cases of amyotrophic lateral sclerosis (ALS), yet its underlying genomic mechanisms remain unclear. Using mouse embryonic stem cell differentiation, we show that CREST is essential for motor neuron maturation by activating mature neuronal genes and repressing progenitor genes.

Mechanistically, CREST interacts with the neuronal transcription factors ONECUT1 and NGN2, and the BRM ATPase to target pre-existing accessible enhancers of neuronal genes. CREST deletion selectively prevents CBP histone acetyltransferase recruitment, reducing histone H3K27 acetylation at these enhancers and downregulating target gene expression, without disrupting BRM binding, enhancer accessibility, or long-range chromatin architecture. Overall, our findings identify CREST as a crucial epigenetic mediator bridging lineage-specific transcription factors, BAF complexes, and CBP to activate enhancers and promote proper motor neuron development.

We thank Andy Lu, Ravneet Jaura, Kavindu Puwakdandawa, and other members of the Rhee laboratory for their experimental support and valuable discussions. We thank Miguel Ramalho-Santos, Jennifer Mitchell, Tim Westwood (University of Toronto), Frank Pugh (Cornell University), and the Transcription, Chromatin and Epigenetics (TranCE) Community in Toronto for valuable comments. Alexandrova (SickKids Hospital) for computational assistance.

We also thank the Genome Quebec Centre of Expertise and Services and the Centre for Applied Genomics at The Hospital for Sick Children for performing the NGS sequencing. This work was supported by funding from Natural Sciences and Engineering Research Council of Canada (NSERC) grant RGPIN-2018-0604, John Evans Leader Fund grant CFI37547, Connaught Fund grant NR504899, and Canadian Institutes of Health Research (CIHR) project grant 202203PJT-183582 to H.S.R. A.J.L. and I.J. were supported by the Ministry of Science and Information and Communication Technology (ICT) through the National Research Foundation in Republic of Korea (RS-2026-25475699, RS-2025-02214225, RS-2023-00265820, N10260004).

These authors contributed equally: Kaitlin N. Department of Cell & Systems Biology, University of Toronto, Toronto, ON, Canada Kaitlin N. Montanera, Alyssa Ialongo Del Prete, Ssu-Yu Yeh & Ho Sung Rhee Department of Biology, University of Toronto, Mississauga, ON, Canada Kaitlin N.

Montanera, Woobin Yun, Alyssa Ialongo Del Prete, Ssu-Yu Yeh & Ho Sung Rhee Department of Biological Sciences, Korea Advanced Institute of Science and Technology (KAIST), Daejeon, Republic of Korea KAIST InnoCORE AI-CRED Institute, Korea Advanced Institute of Science and Technology (KAIST), Daejeon, Republic of Korea Correspondence to Inkyung Jung or Ho Sung Rhee. The authors declare no competing interests. Publisher’s note Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.