Disordered regions and folded modules in CAF-1 promote histone deposition in S. pombe

Ouasti, F.; Audin, M.; Freon, K.; Quivy, J.-P.; Tachekort, M.; Cesard, E.; Thureau, A.; Ropars, V.; Varela, P. F.; Moal, G.; Soumana-Amadou, I.; Uryga, A.; Legrand, P.; Andreani, J.; Guerois, R.; Almouzni, G.; lambert, s. A.; Ochsenbein, F.

doi:10.1101/2023.06.02.543505

bioRxiv · 10.1101/2023.06.02.543505

Disordered regions and folded modules in CAF-1 promote histone deposition in S. pombe

Ouasti, F.·Audin, M.·Freon, K.·Quivy, J.-P.·Tachekort, M.·Cesard, E.·Thureau, A.·Ropars, V.·Varela, P. F.·Moal, G.·Soumana-Amadou, I.·Uryga, A.·Legrand, P.·Andreani, J.·Guerois, R.·Almouzni, G.·lambert, s. A.·Ochsenbein, F.

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Abstract

Genome and epigenome integrity in eukaryotes depends on the proper coupling of histone deposition with DNA synthesis. This process relies on the evolutionary conserved histone chaperone CAF-1 for which the links between structure and functions are still a puzzle. While studies of the S. cerevisiae CAF-1 complex enabled to propose a model for the histone deposition mechanism, we still lack a framework to demonstrate its generality and in particular, how its interaction with the polymerase accessory factor PCNA is operating. Here, we reconstituted a complete SpCAF-1 from fission yeast. We characterized its dynamic structure using NMR, SAXS and molecular modeling together with in vitro and in vivo functional studies on rationally designed interaction mutants. Importantly, we identify the unfolded nature of the acidic domain which folds up when binding to histones. We also show how the long KER helix mediates DNA binding and stimulates SpCAF-1 association with PCNA. Our study highlights how the organization of CAF-1 comprising both disordered regions and folded modules enables the dynamics of multiple interactions to promote synthesis-coupled histone deposition essential for its DNA replication, heterochromatin maintenance, and genome stability functions.

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Ouasti, F., Audin, M., Freon, K., Quivy, J.-P., Tachekort, M., Cesard, E., Thureau, A., Ropars, V., Varela, P. F., Moal, G., Soumana-Amadou, I., Uryga, A., Legrand, P., Andreani, J., Guerois, R., Almouzni, G., lambert, s. A., Ochsenbein, F.. 2023-06-03. Disordered regions and folded modules in CAF-1 promote histone deposition in S. pombe. https://doi.org/10.1101/2023.06.02.543505

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