A Rahman Syndrome mutation in histone H1.4 disrupts chromatin compaction and phase separation
- Ramachandran Boopathi
- Isabel Garcia-Saez
- Serhan Turunç
- Imtiaz Nisar Lone
- Ashok Kumar
- Abed Alkarem Abu Alhaija
- Jeffrey J. Hayes
- Jan Bednar
- Muhammed Kasim Diril
- Dimitar Iliev
- Anastas Gospodinov
- Aline Le Roy
- Dimitrios Skoufias
- Dimitar Angelov
- Ali Hamiche
- Seyit Kale
- Stefan Dimitrov
- Carlo Petosa
2026-05-22
Rahman syndrome is a rare developmental disorder caused by frameshift mutations in linker histone H1.4 that produce a truncated carboxy-terminal domain with reduced positive charge. We investigated the effects of a disease-associated mutation on chromatin structure and dynamics, focusing on H1.4-bound nucleosomes and hexanucleosomal arrays. We report that this mutation induces a more extended and flexible array conformation, characterized by enhanced linker DNA accessibility and an inability to form compact, regularly stacked nucleosome structures. Notably, mutant H1.4-bound arrays show a reduced capacity to undergo liquid-liquid and liquid-solid phase separation, closely resembling linker histone-free arrays. Molecular dynamics simulations corroborated by fluorescence resonance energy transfer measurements indicate that the mutated carboxy-terminal domain interacts with a shorter linker DNA segment, resulting in a more open nucleosome conformation. Consistent with these structural changes, the mutation significantly enhances H1.4 mobility within cell nuclei, reflecting a weaker chromatin association. The combined data suggest that Rahman syndrome-associated mutations promote an aberrantly relaxed chromatin state, potentially leading to the dysregulation of gene expression that may drive disease pathology. These findings underscore the essential role of the carboxy-terminal domain in chromatin compaction and provide mechanistic insights into the molecular etiology of Rahman syndrome.