bims-crepig Biomed News
on Chromatin regulation and epigenetics in cell fate and cancer
Issue of 2025–07–13
27 papers selected by
Connor Rogerson, University of Cambridge



  1. Nucleic Acids Res. 2025 Jul 08. pii: gkaf607. [Epub ahead of print]53(13):
      To date, most studies explored changes in 3D-genome organization between different tissues or during differentiation, which involve massive reprogramming of transcriptional programs. Much fewer studies examined alterations in genome organization in response to cellular stress, which involves less pervasive transcriptional modulation. Here, we examined associations between spatial chromatin organization and gene expression in two different biological contexts: transcriptional programs determining cell identity and transcriptional responses to stress, using p53 activation as a model. We selected 10 cell lines of diverse tissues, and in each performed micro-C, RNA-seq, and p53 ChIP-seq, before and after p53 induction. In the comparison between cell types, we delineated marked correlations between gene expression and spatial genome organization and identified hundreds of active enhancer-promoter loops associated with the expression of cell-type marker genes. In contrast, within each cell type, no such links were observed for expression changes induced by p53 activation, even for enhancers and promoters activated by p53 binding. Our analysis points to a fundamental difference between chromatin interactions that define cell identity and those that are established in response to cellular stress. Our results on p53-induced transcriptional responses support the recently proposed TF activity gradient model, which speculated a contact-independent mechanism for enhancer-promoter communication.
    DOI:  https://doi.org/10.1093/nar/gkaf607
  2. PLoS Biol. 2025 Jul;23(7): e3003239
      After egg fertilization, an initially silent embryonic genome is transcriptionally activated during the maternal-to-zygotic transition. In zebrafish, maternal vertebrate pluripotency factors Nanog, Pou5f3 (OCT4 homolog), and Sox19b (SOX2 homolog) (NPS) play essential roles in orchestrating embryonic genome activation, acting as "pioneers" that open condensed chromatin and mediate acquisition of activating histone modifications. However, some embryonic gene transcription still occurs in the absence of these factors, suggesting the existence of other mechanisms regulating genome activation. To identify chromatin signatures of these unknown pathways, we profiled the histone modification landscape of zebrafish embryos using CUT&RUN. Our regulatory map revealed two subclasses of enhancers distinguished by presence or absence of H3K4me2. Enhancers lacking H3K4me2 tend to require NPS factors for de novo activation, while enhancers bearing H3K4me2 are epigenetically bookmarked by DNA hypomethylation to recapitulate gamete activity in the embryo, independent of NPS pioneering. Thus, parallel enhancer activation pathways combine to induce transcriptional reprogramming to pluripotency in the early embryo.
    DOI:  https://doi.org/10.1371/journal.pbio.3003239
  3. Nucleic Acids Res. 2025 Jul 08. pii: gkaf598. [Epub ahead of print]53(13):
      Transcription initiation by RNA polymerase II is facilitated by coactivators that recruit general initiation factors to promoters. Coactivator complexes transcription factor IID (TFIID) and Spt-Ada-Gcn5-acetyltransferase (SAGA) recruit TATA-binding protein (TBP), while SAGA also enhances transcription by histone acetylation via the HAT Gcn5. It was proposed that most yeast genes depend exclusively on TFIID, with only ∼10% requiring cumulative contributions by SAGA and TFIID for efficient transcription. It was further suggested that genes induced by Gcn4, transcriptional activator of amino acid biosynthetic genes induced by amino acid starvation, depend on the HAT but not the TBP-recruitment function of SAGA. At odds with this model, ChIP-sequencing of TBP and Pol II subunit Rpb1 revealed that deleting SPT3 or SPT8, but not GCN5, reduced TBP binding at many Gcn4 target genes. In contrast, deleting GCN5 but not SPT3 or SPT8 impaired promoter histone eviction at the highly remodeled subset of induced genes, whereas transcription was broadly reduced by all three SAGA mutations. Nuclear depletion of TFIID subunit Taf1 generally reduced TBP recruitment at these and most other SAGA-dependent genes only in cells lacking Spt3 or Spt8. We conclude that SAGA is crucial for TBP recruitment via Spt3/Spt8, beyond its role in histone acetylation, and functions non-redundantly with TFIID in the Gcn4 transcriptome of amino acid-starved cells.
    DOI:  https://doi.org/10.1093/nar/gkaf598
  4. Nat Struct Mol Biol. 2025 Jul 07.
      Transcription poses a major challenge for genome stability. The RECQL5 helicase helps safeguard genome integrity and is the only member of the human RecQ helicase family that directly binds to RNA polymerase II (Pol II) and affects its progression. While RECQL5 mitigates transcription stress in cells, the molecular mechanism underlying this phenomenon is unclear. Here, we use cryo-electron microscopy to determine the structures of stalled human Pol II elongation complexes (ECs) bound to RECQL5. Our structures reveal the molecular interactions stabilizing RECQL5 binding to the Pol II EC and highlight its role as a transcriptional roadblock. Additionally, we find that, in its nucleotide-free state, RECQL5 twists the downstream DNA in the EC and, upon nucleotide binding, undergoes a conformational change that allosterically induces Pol II toward a post-translocation state. We propose that this mechanism may help restart Pol II elongation and, therefore, contribute to reducing transcription stress.
    DOI:  https://doi.org/10.1038/s41594-025-01611-8
  5. Nat Cell Biol. 2025 Jul 10.
      The regulation of gene expression is governed at multiple levels of chromatin organization. However, how gene regulation is co-ordinated remains relatively unexplored. Here we develop Dam&ChIC, a method that enables retrospective and multifactorial chromatin profiling in single cells. Dam&ChIC employs chromatin labelling in living cells with m6A to acquire a past chromatin state, coupled with an antibody-mediated readout to capture the present chromatin state. Analyses of diverse factor combinations highlight its versatility and superior resolution. By tracking lamina-associated domain inheritance over the cell cycle, we showcase that Dam&ChIC provides retrospective single-cell chromatin data. When applied in random X chromosome inactivation, Dam&ChIC disentangles the temporal order of chromatin remodelling events. Upon mitotic exit and following Xist expression, the inactive X chromosome undergoes extensive genome-lamina detachment, preceding spreading of Polycomb. We anticipate that Dam&ChIC will be instrumental in unravelling the interconnectivity and order of gene-regulatory events underlying cell-state changes during development.
    DOI:  https://doi.org/10.1038/s41556-025-01687-w
  6. Commun Biol. 2025 Jul 07. 8(1): 1016
      The FOX transcription factor family plays a pivotal role in the malignant progression of tumors. We propose a hypothesis that FOXQ1 recruits p300 and BRD4 to super-enhancer regions. Our findings indicate that p300 acetylates Lys190 of FOXQ1, resulting in its recognition and binding by BRD4. Subsequently, BRD4 recruits RNA-Pol II to form a "FOXQ1-p300-BRD4-RNA Pol II" complex, which then binds to the super-enhancers of target genes. Meanwhile, acetylation at Lys190 of FOXQ1 directly enhances its binding affinity to super-enhancers. Consequently, more target oncogenes can be transcribed to promote CRC proliferation and metastasis. Our results suggest that FOXQ1 acts as a key regulator of super-enhancers, providing insights into its role in CRC and highlighting its potential as a therapeutic target.
    DOI:  https://doi.org/10.1038/s42003-025-08430-z
  7. Nat Neurosci. 2025 Jul 08.
      Diverse types of GABAergic projection neuron and interneurons of the telencephalon derive from progenitors in a ventral germinal zone called the ganglionic eminence. Using single-cell transcriptomics, chromatin accessibility profiling, lineage tracing, birthdating, transplantation across developmental stages and perturbation sequencing in mouse embryos, we investigated how progenitor competence influences the maturation and differentiation of these neurons. We found that the temporal progression of neurogenesis shapes maturation competence in ganglionic eminence progenitors, influencing how their progeny progress toward mature states. By contrast, differentiation competence-defined as the ability of progenitors to produce diverse transcriptomic identities-was maintained throughout neurogenesis. Chromatin remodeling, together with a regulatory module composed of the transcription factor NFIB and its target genes, influenced maturation competence in late-born neurons. These findings reveal how transcriptional programs and chromatin accessibility govern neuronal maturation and the diversification of GABAergic neuron subtypes during neurodevelopment.
    DOI:  https://doi.org/10.1038/s41593-025-01999-y
  8. Nat Struct Mol Biol. 2025 Jul 07.
      Abnormally fast transcription elongation can lead to detrimental consequences such as transcription-replication collisions, altered alternative splicing patterns and genome instability. Therefore, elongating RNA polymerase II (Pol II) requires mechanisms to slow its progression, yet the molecular basis of transcription braking remains unclear. RECQL5 is a DNA helicase that functions as a general elongation factor by slowing down Pol II. Here we report cryo-electron microscopy structures of human RECQL5 bound to multiple transcription elongation complexes. Combined with biochemical analysis, we identify an α-helix of RECQL5 responsible for binding Pol II and slowdown of transcription elongation. We further reveal that the transcription-coupled DNA repair (TCR) complex allows Pol II to overcome RECQL5-induced transcription braking through concerted actions of its translocase activity and competition with RECQL5 for engaging Pol II. Additionally, RECQL5 inhibits TCR-mediated Pol II ubiquitination to prevent activation of the DNA repair pathway. Our results suggest a model in which RECQL5 and the TCR complex coordinately regulate transcription elongation rates to ensure transcription efficiency while maintaining genome stability.
    DOI:  https://doi.org/10.1038/s41594-025-01586-6
  9. Genome Biol. 2025 Jul 07. 26(1): 196
      Single-cell profiling protein-chromatin interactions is often constrained by complex workflows, high cost, or dependence on specialized equipment. We present indexed tagmentation-based single-cell CUT&Tag-sequencing (IT-scC&T-seq), a modular, plate-based strategy using three-round combinatorial barcoding. IT-scC&T-seq robustly profiles histone modifications and transcription factors with high specificity and throughput, supporting simultaneous analysis of multiple samples and epitopes. Notably, it enables sensitive single-cell mapping of lamina-associated domains, low-abundance chromatin features previously difficult to resolve. Applied to adult mouse mammary gland, the method reveals cell-type-specific chromatin landscapes and lineage-regulatory dynamics. Together, IT-scC&T-seq provides a scalable, cost-effective, and broadly accessible approach for high-resolution chromatin profiling.
    Keywords:  CUT&Tag; Epigenetics; Histone modification; Indexed tagmentation; Lamina-associated domains (LADs); Mammary gland development; Single-cell omics
    DOI:  https://doi.org/10.1186/s13059-025-03661-z
  10. PLoS Genet. 2025 Jul 11. 21(7): e1011778
      Oligodendrocytes (OLs) assemble myelin sheaths around axons in central nervous system (CNS). Myelin is essential for the saltatory conduction of action potentials and also performs other critical functions for the operation of the CNS. Sox10 (SRY-box containing gene 10) is a high-mobility group transcription factor that orchestrates the development of OLs. Despite its key role in OL biology, there is scant information on how the expression of Sox10 is regulated in OL lineage cells. Especially, OL enhancers that control its transcription remain elusive. We have recently developed an innovative method that rationally links OL enhancers to target genes. This study applied the new method to Sox10, uncovering two OL enhancers for it (termed Sox10-E1 and Sox10-E2). Epigenome editing analysis revealed that Sox10-E1 and Sox10-E2 regulate Sox10 expression non-redundantly. Luciferase assay and human and mouse brain multi-omics data show that, during the differentiation of OL precursor cells (OPCs) into OLs, the enhancer activity of Sox10-E1 does not change while that of Sox10-E2 decreases significantly. Chromatin interaction data indicate that the two Sox10 enhancers lie close to the border of the Sox10 topologically associating domain (TAD). Consistently, Pick1, a gene that is near the Sox10 TAD border, is also under the transcriptional control of Sox10-E1 and Sox10-E2. Hence, genomic deletions involving Sox10-E1 and Sox10-E2 would perturb not only SOX10, but also PICK1 and other genes, and may cause a pathology that is more complex than that of conventional Waardenburg-Shah syndrome that results from SOX10 coding mutations.
    DOI:  https://doi.org/10.1371/journal.pgen.1011778
  11. Biosystems. 2025 Jul 07. pii: S0303-2647(25)00141-8. [Epub ahead of print]254 105531
      Identifying the regulatory relationships between transcription factors and target genes is fundamental to understanding molecular regulatory mechanisms in biological processes including development and disease occurrence. Therefore, resolving the relationships between cis-regulatory elements and genes using single-cell multi-omics data is important for understanding transcriptional regulation. Here, scSAGRN is proposed as a framework for inferring gene regulatory networks from single-cell multi-omics. scSAGRN incorporates spatial association to compute correlations between gene expression and chromatin openness data, connects distal cis-regulatory elements to genes, infers gene regulatory networks and identifies key transcription factors. The approach is benchmarked using real single-cell datasets, and scSAGRN shows superior performance in TF recovery, peak-gene linkage prediction, and TF-gene linkage prediction compared to existing methods. Meanwhile, in human peripheral blood mononuclear cells dataset, mouse cerebral cortex dataset and mouse embryonic brain cells dataset, scSAGRN demonstrates its capability to infer gene regulatory networks and identify transcription factors. Overall, scSAGRN provides a reference for predicting transcriptional regulatory patterns from single-cell multi-omics data.
    Keywords:  Cis-regulatory element (CRE); Gene regulatory network (GRN); Single-cell multi-omics; Transcription factor (TF); Weighted nearest neighbor (WNN)
    DOI:  https://doi.org/10.1016/j.biosystems.2025.105531
  12. Nat Genet. 2025 Jul 09.
      Transposable elements (TEs) are scattered across mammalian genomes. Silencing of TEs prevents harmful effects caused by either global activation leading to genome instability or insertional mutations disturbing gene transcription. However, whether the activation of a TE can cause disease without directly affecting gene expression is largely unknown. Here we show that a TE insertion can adopt nearby regulatory activity, resulting in the production of cell-type-specific viral-like particles (VLPs) that affect embryo formation. Failure to silence an LTR retrotransposon inserted upstream of the Fgf8 gene results in their co-expression during mouse development. VLP assembly in the Fgf8-expressing cells of the developing limb triggers apoptotic cell death, resulting in a limb malformation resembling human ectrodactyly. The phenotype can be rescued by mutating the retrotransposon coding sequence, thus preventing its full endogenous retroviral cycle. Our findings illustrate that TE insertions can be incorporated into the local genomic regulatory landscape and that VLP production in post-implantation embryos can cause developmental defects.
    DOI:  https://doi.org/10.1038/s41588-025-02248-5
  13. Nucleic Acids Res. 2025 Jul 08. pii: gkaf612. [Epub ahead of print]53(13):
      Post-translational modifications of histone tails constitute a key epigenetic mechanism controlling chromatin environment and gene transcription. Serotonylation of histone H3Q5 (H3Q5ser) is a recently discovered mark associated with active transcription of RNA polymerase II (pol II)-transcribed genes. The direct link between H3Q5ser and the pol II transcription machinery relies on the TFIID subunit TAF3. The presence of H3Q5ser enhances TAF3 binding to H3K4me3, but the molecular determinants underlying this interaction remained unclear. Here, we resolve the binding mode of TAF3-PHD to H3K4me3Q5ser identifying a novel binding surface for H3Q5ser using solution nuclear magnetic resonance spectroscopy. This reveals how H3Q5ser recognizes a conserved surface of the TAF3-PHD via CH-π interactions in an edge-face conformation involving a proline residue stabilized by a tryptophan. This combination of proline and tryptophan is unique to the PHD finger of TAF3 and conserved among TAF3 orthologues. Our findings establish a framework for the molecular recognition of serotonylated chromatin, laying the foundation for developing epigenetic inhibitors targeting serotonylation-dependent transcriptional regulation in neuronal development.
    DOI:  https://doi.org/10.1093/nar/gkaf612
  14. Nat Commun. 2025 Jul 09. 16(1): 6315
      Cellular chromatin displays heterogeneous structure and dynamics, properties that control diverse nuclear processes. Models invoke phase separation of conformational ensembles of chromatin fibers as a mechanism regulating chromatin organization in vivo. Here we combine biochemistry and molecular dynamics simulations to examine, at single base-pair resolution, how nucleosome spacing controls chromatin phase separation. We show that as DNA linkers extend from 25 bp to 30 bp, as exemplars of 10 N + 5 and 10 N (integer N) bp lengths, chromatin condensates become less thermodynamically stable and nucleosome mobility increases. Simulations reveal that this is due to trade-offs between inter- and intramolecular nucleosome stacking, favored by rigid 10 N + 5 and 10 N bp linkers, respectively. A remodeler can induce or inhibit phase separation by moving nucleosomes, changing the balance between intra- and intermolecular stacking. The intrinsic phase separation capacity of chromatin enables fine tuning of compaction and dynamics, likely contributing to heterogeneous chromatin organization in vivo.
    DOI:  https://doi.org/10.1038/s41467-025-61482-x
  15. Nat Commun. 2025 Jul 08. 16(1): 6273
      DNA methylation patterns at crucial short sequence features, such as enhancers and promoters, may convey key information about cell lineage and state. The need for high-resolution single-cell DNA methylation profiling has therefore become increasingly apparent. Existing single-cell whole-genome bisulfite sequencing (scWGBS) studies have both methodological and analytical shortcomings. Inefficient library generation and low CpG coverage mostly preclude direct cell-to-cell comparisons and necessitate the use of cluster-based analyses, imputation of methylation states, or averaging of DNA methylation measurements across large genomic bins. Such summarization methods obscure the interpretation of methylation states at individual regulatory elements and limit our ability to discern important cell-to-cell differences. We report an improved scWGBS method, single-cell Deep and Efficient Epigenomic Profiling of methyl-C (scDEEP-mC), which offers efficient generation of high-coverage libraries. scDEEP-mC allows for cell type identification, genome-wide profiling of hemi-methylation, and allele-resolved analysis of X-inactivation epigenetics in single cells. Furthermore, we combine methylation and copy-number data from scDEEP-mC to identify single, actively replicating cells and profile DNA methylation maintenance dynamics during and after DNA replication. These analyses unlock further avenues for exploring DNA methylation regulation and dynamics and illustrate the power of high-complexity, highly efficient scWGBS library construction as facilitated by scDEEP-mC.
    DOI:  https://doi.org/10.1038/s41467-025-61589-1
  16. Nat Plants. 2025 Jul 09.
      Phosphorylation at the carboxy-terminal domain of the largest subunit of RNA polymerase II plays a critical role in transcription, and histone acetylation is correlated with active transcription. However, the regulatory mechanisms by which histone acetylation modulates RNA polymerase II phosphorylation in plants remain unclear. Here we show that two functionally redundant bromodomain-containing proteins, global transcription factor group E2 (GTE2) and GTE7, can bind to acetylated histone H4. Both GTE2 and GTE7 interact with cyclin-dependent kinase-like 9 (CDKL9), which belongs to a plant-specific CDKL group. Unlike canonical CDKs, CDKL9 functions in a cyclin- and CDK-activating-kinase-independent manner and can phosphorylate at least the serine 2 and serine 5 residues of the carboxy-terminal domain in vitro. The GTE2/GTE7-CDKL9 complex is required to maintain serine 2 and serine 5 phosphorylation under heat stress. Consistently, loss-of-function gte2/gte7 and cdkl9 mutants show similar heat-sensitive phenotypes. We also demonstrate that the acetylated-histone-binding activity of GTE7 is essential for the association of CDKL9 with chromatin and for plant heat tolerance. Together, these findings provide mechanistic insight into transcriptional regulation via histone acetylation in response to heat stress and suggest that plants might have evolved a unique group of carboxy-terminal domain kinases for stress tolerance.
    DOI:  https://doi.org/10.1038/s41477-025-02044-3
  17. Nat Commun. 2025 Jul 05. 16(1): 6206
      Integrator-PP2A (INTAC) is a highly modular complex orchestrating the transition of paused RNA polymerase II into productive elongation or promoter-proximal premature termination, with its loss resulting in transcription dysregulation and genome instability. Here, we identify human DSS1-a flexible 70-residue protein found in multiple functionally diverse complexes including the 26S proteasome-as an integral subunit of the INTAC backbone. Structural analysis of DSS1-INTAC, both alone and in association with paused polymerase, demonstrates intimate interactions between DSS1 and the INTAC backbone. We identify tryptophan 39 of DSS1 as being critical for interacting with INTAC and find that its mutation disrupts DSS1's interaction with INTAC, while maintaining DSS1's interaction with the proteasome. This substitution not only impairs INTAC-dependent transcriptional regulation, but also reveals that INTAC is DSS1's major chromatin-bound form. Together, our findings reveal a role for DSS1 in supporting the structure and regulatory functions of INTAC.
    DOI:  https://doi.org/10.1038/s41467-025-61257-4
  18. Nucleic Acids Res. 2025 Jul 08. pii: gkaf640. [Epub ahead of print]53(13):
      Single-cell long-read concatemer sequencing (scNanoHi-C) technology provides unique insights into the higher-order chromatin structure across the genome in individual cells, crucial for understanding 3D genome organization. However, the lack of specialized analytical tools for scNanoHi-C data impedes progress, as existing methods, which primarily focus on scHi-C technologies, do not fully address the specific challenges of scNanoHi-C, such as sparsity, cell-specific variability, and complex chromatin interaction networks. Here, we introduce DeepNanoHi-C, a novel deep learning framework specifically designed for scNanoHi-C data, which leverages a multistep autoencoder and a Sparse Gated Mixture of Experts (SGMoE) to accurately predict chromatin interactions by imputing sparse contact maps, thereby capturing cell-specific structural features. DeepNanoHi-C effectively captures complex global chromatin contact patterns through the multistep autoencoder and dynamically selects the most appropriate expert from a pool of experts based on distinct chromatin contact patterns. Furthermore, DeepNanoHi-C integrates multiscale predictions through a dual-channel prediction net, refining complex interaction information and facilitating comprehensive downstream analyses of chromatin architecture. Experimental validation shows that DeepNanoHi-C outperforms existing methods in distinguishing cell types and demonstrates robust performance in data imputation tasks. Additionally, the framework identifies single-cell 3D genome features, such as cell-specific topologically associating domain (TAD) boundaries, further confirming its ability to accurately model chromatin interactions. Beyond single-cell analysis, DeepNanoHi-C also uncovers conserved genomic structures across species, providing insights into the evolutionary conservation of chromatin organization.
    DOI:  https://doi.org/10.1093/nar/gkaf640
  19. Gastroenterology. 2025 Jul 07. pii: S0016-5085(25)05732-4. [Epub ahead of print]
       BACKGROUND AND AIMS: Aberrant epigenetic programs that suppress differentiation and enhance plasticity drive colorectal cancer (CRC), yet the molecular determinants underlying these processes remain elusive. We aimed to identify and characterize epigenetic regulators of CRC differentiation, uncovering mechanisms that reprogram cancer cell states.
    METHODS: A small molecule library targeting epigenetic regulators was screened using an endogenous dual reporter system. We evaluated lead compounds in mouse and human CRC models via histopathology, cellular assays, epigenetic studies, mass-spectrometry-based histone modification profiling, and single cell RNA-sequencing. Integrative analyses of drug-induced chromatin dynamics, gene expression, target engagement, and histone marks elucidated molecular mechanisms. Focused genetic screens were conducted to identify regulators of HDAC1/2-mediated differentiation.
    RESULTS: We found that inhibition of histone deacetylase (HDAC) 1/2 catalytic domain promotes CRC differentiation and suppresses tumor growth. Unbiased profiling of histone modifications identified H3K27ac and H3K9ac as critical regulatory marks, with genome-wide analyses demonstrating their enrichment at HDAC1/2-bound regions associated with open chromatin and upregulated differentiation genes. Disrupting H3K27ac by targeted degradation of acetyltransferase EP300 reversed the differentiation phenotype induced by HDAC1/2 inhibition in a patient-derived CRC organoid. Genetic screens revealed that DAPK3 contributes to H3K27ac-mediated CRC differentiation induced by HDAC1/2 inhibition.
    CONCLUSIONS: Our findings establish histone acetylation as a chemically targetable mechanism governing CRC cell fate and demonstrate that epigenetic reprogramming can be leveraged as a therapeutic strategy. By identifying HDAC1/2 inhibition as a driver of differentiation and revealing H3K27ac as a key regulatory mark, this study provides a framework for targeting chromatin-modifying enzymes to counteract CRC plasticity and improve treatment outcomes.
    Keywords:  colorectal cancer; epigenetic regulation; intestinal differentiation; stem cell
    DOI:  https://doi.org/10.1053/j.gastro.2025.07.003
  20. Biophys J. 2025 Jul 03. pii: S0006-3495(25)00419-9. [Epub ahead of print]
      Many transcription factors regulate DNA accessibility and gene expression by recognizing post-translational modifications on histone tails within nucleosomes. These interactions are often studied in vitro using short peptide mimics of histone tails, which may overlook conformational changes that occur in the full nucleosomal context. Here, we employ molecular dynamics simulations to investigate the binding dynamics of the PHD finger and bromodomain of BPTF, both in solution and bound to either a histone H3 peptide or a full nucleosome. Our results show that BPTF adopts distinct conformational states depending on its binding context, with nucleosome engagement inducing compaction of the multidomain structure. PHD finger binding displaces the H3 tail from DNA, increasing H3 tail flexibility while promoting compensatory binding of the H4 tail to nucleosomal DNA. This redistribution of histone-DNA contacts weakens overall hydrogen bonding with DNA, suggesting localized destabilization of the nucleosome core. Despite electrostatic repulsion limiting direct reader-DNA contacts, strong Van der Waals interactions with the H3 tail stabilize binding. Our results provide atomistic insight into how BPTF engagement modulates nucleosome structure and may facilitate chromatin remodeling.
    DOI:  https://doi.org/10.1016/j.bpj.2025.06.042
  21. Nat Commun. 2025 Jul 10. 16(1): 6375
      Heterochromatin protein 1 (HP1) is a multifunctional chromatin-associated protein conserved from fission yeast to mammals. HP1 has been suggested to drive heterochromatin formation via phase separation. However, there is seemingly conflicting evidence about HP1 phase-separating in different systems or not. Here, we assess the phase separation behavior of HP1 from fission yeast, fruit fly and mouse in vitro and in mammalian cells side-by-side. We find that HP1 from fission yeast and fly can undergo liquid-liquid phase separation and induce heterochromatin coalescence in mouse cells, in stark contrast to HP1 from mouse. Induced heterochromatin coalescence has only mild effects on gene expression. We link the decreasing phase separation propensity of HP1 homologs to their decreasing intrinsic disorder and their increasing sensitivity to HP1 paralogs antagonizing phase separation. Our work elucidates the relationship between phase separation, nuclear organization and gene expression, and highlights the evolutionary dimension of protein phase separation control.
    DOI:  https://doi.org/10.1038/s41467-025-61749-3
  22. Nat Commun. 2025 Jul 08. 16(1): 6288
      Dishevelled (DVL) is a crucial component of the Wnt-signaling pathway and is vital for multiple physiological processes. Previously thought to have a classically cytoplasmic role, the discovery of DVL nuclear translocation reframed how it is viewed functionally. Although significant progress has been made in understanding the nuclear functions of DVL, further research is required to clarify its roles in transcriptional and epigenetic regulation. A key unresolved question is whether nuclear DVL1 associates with a transcription factor partner. We show here that modulation of DVL1 expression globally affects the transcriptomic landscape. Additionally, analysis of DVL1 ChIP-sequencing allowed us to map genome-wide binding sites, revealing the extensive reach of DVL1 binding. Integration of RNA-sequencing and ChIP-sequencing further revealed ETS1 as a transcription factor binding partner which targets nuclear DVL1 to specific genomic loci. These findings provide insight into the contribution of DVL1 in transcription and clarify aspects of its elusive nuclear function.
    DOI:  https://doi.org/10.1038/s41467-025-61551-1
  23. Nat Methods. 2025 Jul 10.
      Epigenetic modifications on natural chromosomes are inherited and maintained in a default state, making it challenging to remove intrinsic marks to study the fundamental principles of their establishment and further influence on transcriptional regulation. In this study, we developed SynNICE, a method for assembling and delivering intact, naive, synthetic megabase (Mb)-scale human DNA into early mouse embryos, to study de novo epigenetic regulation. By assembling and delivering a 1.14-Mb human AZFa (hAZFa) locus, we observed the spontaneous incorporation of murine histones and the establishment of DNA methylation at the one-cell stage. Notably, DNA methylation from scratch strongly enriches at repeat sequences without H3K9me3 reinforcement. Furthermore, the transcription of hAZFa initiated at the four-cell stage is regulated by newly established DNA methylation. This method provides a unique platform for exploring de novo epigenomic regulation mechanisms in higher animals.
    DOI:  https://doi.org/10.1038/s41592-025-02746-8
  24. Genome Res. 2025 Jul 08.
      In mammalian interphase cells, genomes are folded by cohesin loop extrusion limited by directional CTCF barriers. This process enriches cohesin at barriers, isolates neighboring topologically associating domains, and elevates contact frequency between convergent CTCF barriers across the genome. However, recent in vivo measurements present a puzzle: reported CTCF residence times on chromatin are in the range of a few minutes, whereas cohesin lifetimes are much longer. Can the observed features of genome folding result from relatively transient barriers? To address this question, we develop a dynamic barrier model, where CTCF sites switch between bound and unbound states. Using this model, we investigate how barrier dynamics would impact observables for a range of experimental genomic and imaging data sets, including ChIP-seq, Hi-C, and microscopy. We find the interplay of CTCF and cohesin binding timescales influence the strength of each of these features, leaving a signature of barrier dynamics even in the population-averaged snapshots offered by genomic data sets. First, in addition to barrier occupancy, barrier bound times are crucial for instructing features of genome folding. Second, the ratio of boundary to extruder lifetime greatly alters simulated ChIP-seq and simulated Hi-C. Third, large-scale changes in chromosome morphology observed experimentally after increasing extruder lifetime require dynamic barriers. By integrating multiple sources of experimental data, our biophysical model argues that CTCF barrier bound times effectively approach those of cohesin extruder lifetimes. Together, we demonstrate how models that are informed by biophysically measured protein dynamics broaden our understanding of genome folding.
    DOI:  https://doi.org/10.1101/gr.280108.124
  25. Science. 2025 Jul 10. 389(6756): eadn6121
      Human neurons programmed through transcription factor (TF) overexpression model neuronal differentiation and disease. However, the diversity of neuronal subtypes programmable in vitro remains unresolved. We modulated developmental signaling pathways combined with TF overexpression to explore the spectrum of neuron subtypes generated from pluripotent stem cells. We screened 480 morphogen signaling modulations coupled with TF induction using a multiplexed single-cell transcriptomic readout. Analysis of 700,000 cells identified diverse excitatory and inhibitory neurons patterned along the developmental axes of the neural tube. Patterning neural progenitors prior to TF overexpression expanded neuronal diversity by enabling access to regulons active in primary tissue counterparts. Our approach provides a strategy for programming diverse human cell subtypes as well as investigating how cooperative signaling drives neuronal fate.
    DOI:  https://doi.org/10.1126/science.adn6121
  26. Cell Rep. 2025 Jul 09. pii: S2211-1247(25)00720-X. [Epub ahead of print]44(7): 115949
      We have previously shown that the nuclear factor κB (NF-κB) inhibitor IκBα binds chromatin together with polycomb repressor complex 2 (PRC2) to confer responsiveness to PRC2 targets in the presence of inflammatory cues. This alternative function has been elusive in both physiological and disease conditions because of the predominant role of IκBα as a negative regulator of NF-κB. Here, we uniquely characterize the specific residues of IκBα that allow the generation of separation-of-function (SOF) mutants that are defective for either NF-κB-related (SOFΔNF-κB) or chromatin-related (SOFΔH2A,H4) activities. Expression of IκBα SOFΔNF-κB, but not SOFΔH2A/H4, is sufficient to negatively regulate a specific stemness program in intestinal cells, thereby rescuing the differentiation block imposed by IκBα deficiency. Using a chromatin immunoprecipitation (ChIP) assay, we demonstrate that IκBα binds to several stemness genes that are transcriptionally repressed upon IκBα SOFΔNF-κB induction. Our data suggest that SOF mutants provide an exclusive tool for studying IκBα functions in physiology and disease.
    Keywords:  CP: Stem cell research; IκB⍺; NF-κB; differentiation; histone H4; polycomb; separation of function; stemness
    DOI:  https://doi.org/10.1016/j.celrep.2025.115949
  27. Cell. 2025 Jul 01. pii: S0092-8674(25)00686-5. [Epub ahead of print]
      The regulatory sequences of vertebrate genomes remain incompletely understood. To address this, we developed an ultra-throughput, ultra-sensitive single-nucleus assay for transposase-accessible chromatin using sequencing (UUATAC-seq) protocol that enables the construction of chromatin accessibility landscapes for one species in a 1-day experiment. Using UUATAC-seq, we mapped candidate cis-regulatory elements (cCREs) across five representative vertebrate species. Our analysis revealed that genome size differences across species influence the number but not the size of cCREs. We introduced Nvwa cis-regulatory element (NvwaCE), a mega-task deep-learning model designed to interpret cis-regulatory grammar and predict cCRE landscapes directly from genomic sequences with high precision. NvwaCE demonstrated that regulatory grammar is more conserved than nucleotide sequences and that this grammar organizes cCREs into distinct functional modules. Moreover, NvwaCE accurately predicted the effects of synthetic mutations on lineage-specific cCRE function, aligning with causal quantitative trait loci (QTLs) and genome editing results. Together, our study provides a valuable resource for decoding the vertebrate regulatory language.
    Keywords:  NvwaCE; UUATAC-seq; cCRE; chromatin accessibility landscape; deep learning; genome editing; genomics; mutation effect; regulatory sequence; snATAC-seq
    DOI:  https://doi.org/10.1016/j.cell.2025.06.020