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The nucleosome remodeler Chd1 is required for the re-establishment of nucleosome positioning in the wake of transcription elongation by RNA Polymerase II. Previously, we found that Chd1 occupancy on gene bodies depends on the Rtf1 subunit of the Paf1 complex in yeast. Here, we identify an N-terminal region of Rtf1 and the CHCT domain of Chd1 as sufficient for their interaction and demonstrate that this interaction is direct. Mutations that disrupt the Rtf1-Chd1 interaction result in an accumulation of Chd1 at the 5' ends of Chd1-occupied genes, increased cryptic transcription, altered nucleosome positioning, and concordant shifts in histone modification profiles. We show that a homologous region within mouse RTF1 interacts with the CHCT domains of mouse CHD1 and CHD2. This work supports a conserved mechanism for coupling Chd1 family proteins to the transcription elongation complex and identifies a cellular function for a domain within Chd1 about which little is known.
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http://dx.doi.org/10.1093/nar/gkaf816 | DOI Listing |
Nucleic Acids Res
September 2025
Ohio State Biochemistry Graduate Program, The Ohio State University, Columbus, OH 43210, United States.
Nucleosome repositioning is essential for establishing nucleosome-depleted regions to initiate transcription. This process has been extensively studied using structural, biochemical, and single-molecule approaches, which require homogeneously positioned nucleosomes. This is often achieved using the Widom 601 sequence, a highly efficient nucleosome-positioning element (NPE) selected for its unusually strong binding to the H3-H4 histone tetramer.
View Article and Find Full Text PDFCell Rep
September 2025
Institut Curie, UMR3348, CNRS, Université Paris-Saclay, 91401 Orsay, France. Electronic address:
Alternative splicing enables cells to acquire novel phenotypic traits for adaptation to changes in the environment. However, the mechanisms that allow these dynamic changes to occur in a timely and sustained manner remain unknown. Recent investigations unveiled a new regulatory layer important for splicing dynamics and memory: the chromatin.
View Article and Find Full Text PDFNucleic Acids Res
August 2025
Department of Biological Sciences, University of Pittsburgh, Pittsburgh, PA, 15260, United States.
The nucleosome remodeler Chd1 is required for the re-establishment of nucleosome positioning in the wake of transcription elongation by RNA Polymerase II. Previously, we found that Chd1 occupancy on gene bodies depends on the Rtf1 subunit of the Paf1 complex in yeast. Here, we identify an N-terminal region of Rtf1 and the CHCT domain of Chd1 as sufficient for their interaction and demonstrate that this interaction is direct.
View Article and Find Full Text PDFSTAR Protoc
August 2025
BRIC-Centre for DNA Fingerprinting and Diagnostics (CDFD), Hyderabad, Telangana 500039, India. Electronic address:
Chromatin dynamics plays an important role in medical mycology. Here, we present a protocol to identify nucleosome occupancy and positioning in the fungal pathogen Candida glabrata (Nakaseomyces glabratus). This process involves micrococcal nuclease (MNase) digestion of the chromatin of C.
View Article and Find Full Text PDFNat Struct Mol Biol
August 2025
Division of Genetics, Department of Medicine, Brigham and Women's Hospital and Harvard Medical School, Boston, MA, USA.
Cell fates are controlled by 'pioneers', sequence-specific transcription factors (TFs) that bind recognition motifs on nucleosomes ('pioneer binding'). Pioneers occupy a minority of their recognition sequences in the genome, suggesting that the sequence context regulates their binding. Here we developed PIONEAR-seq, a high-throughput biochemical assay to characterize pioneer binding.
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