RIF1 orchestrates a multi-step restoration of post-replicative H3K9me3

Chen, N.; Bandau, S.; Ravindran Nair Pushkala Kumari, R.; Prevo, B.; Sasaki, T.; Web, S.; di Sanzo, S.; Shapira, D.; Völker-Albert, M.; Gilbert, D. M.; Alabert, C.; Buonomo, S. C. B.

doi:10.1101/2025.07.14.664777

bioRxiv · 10.1101/2025.07.14.664777

RIF1 orchestrates a multi-step restoration of post-replicative H3K9me3

Chen, N.·Bandau, S.·Ravindran Nair Pushkala Kumari, R.·Prevo, B.·Sasaki, T.·Web, S.·di Sanzo, S.·Shapira, D.·Völker-Albert, M.·Gilbert, D. M.·Alabert, C.·Buonomo, S. C. B.

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Abstract

DNA replication causes the dilution of parental histones along with their specific post-translational modifications. The kinetics of restoring these marks on newly incorporated histones dictate how quickly genomic domains regain their epigenetic identity after replication. H3K9me3 is restored extremely slowly; the process of reconstitution, to achieve the pre-replication levels, continues throughout the following G1 phase. The molecular mechanisms behind this slow reconstitution are unknown. We show here that RIF1s reassociation with heterochromatin during mitotic exit is required to set up a chromatin environment permissive for histone methyltransferases to resume H3K9me3 deposition. RIF1 facilitates the recruitment of SUV39H1, HP1 and HP1{beta} and is required for the increased tri-methylation of H3K9 that occurs during G1 phase. RIF1 is also indispensable for recruiting Protein Phosphatase 1 (PP1) to heterochromatin, and the interaction between RIF1 and PP1 is essential for the maintenance of H3K9me3 levels. In addition, RIF1-PP1 complex temporally restrains the activity of Aurora kinase at heterochromatin, ensuring that phosphorylation of H3S10 does not precede replication. This creates a time- window permissive for SUV39Hs to initiate the reinstatement of H3K9me3.

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Chen, N., Bandau, S., Ravindran Nair Pushkala Kumari, R., Prevo, B., Sasaki, T., Web, S., di Sanzo, S., Shapira, D., Völker-Albert, M., Gilbert, D. M., Alabert, C., Buonomo, S. C. B.. 2025-07-15. RIF1 orchestrates a multi-step restoration of post-replicative H3K9me3. https://doi.org/10.1101/2025.07.14.664777

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