Print Email Facebook Twitter CAF-1 deposits newly synthesized histones during DNA replication using distinct mechanisms on the leading and lagging strands Title CAF-1 deposits newly synthesized histones during DNA replication using distinct mechanisms on the leading and lagging strands Author Rouillon, Clément (University Medical Center Utrecht) Eckhardt, Bruna V. (University Medical Center Utrecht) Kollenstart, Leonie (University of Copenhagen) Gruss, Fabian (University Medical Center Utrecht) Verkennis, Alexander E.E. (University Medical Center Utrecht) Rondeel, Inge (University Medical Center Utrecht) van Laar, T. (TU Delft BN/Nynke Dekker Lab; Kavli institute of nanoscience Delft) Albanese, Pascal (Universiteit Utrecht) Dekker, N.H. (TU Delft BN/Nynke Dekker Lab; Kavli institute of nanoscience Delft) Date 2023 Abstract During every cell cycle, both the genome and the associated chromatin must be accurately replicated. Chromatin Assembly Factor-1 (CAF-1) is a key regulator of chromatin replication, but how CAF-1 functions in relation to the DNA replication machinery is unknown. Here, we reveal that this crosstalk differs between the leading and lagging strand at replication forks. Using biochemical reconstitutions, we show that DNA and histones promote CAF-1 recruitment to its binding partner PCNA and reveal that two CAF-1 complexes are required for efficient nucleosome assembly under these conditions. Remarkably, in the context of the replisome, CAF-1 competes with the leading strand DNA polymerase epsilon (Polϵ) for PCNA binding. However, CAF-1 does not affect the activity of the lagging strand DNA polymerase Delta (Polδ). Yet, in cells, CAF-1 deposits newly synthesized histones equally on both daughter strands. Thus, on the leading strand, chromatin assembly by CAF-1 cannot occur simultaneously to DNA synthesis, while on the lagging strand these processes may be coupled. We propose that these differences may facilitate distinct parental histone recycling mechanisms and accommodate the inherent asymmetry of DNA replication. To reference this document use: http://resolver.tudelft.nl/uuid:9371e760-5c94-4b8c-9912-db2d12963000 DOI https://doi.org/10.1093/nar/gkad171 ISSN 0305-1048 Source Nucleic acids research, 51 (8), 3770-3792 Part of collection Institutional Repository Document type journal article Rights © 2023 Clément Rouillon, Bruna V. Eckhardt, Leonie Kollenstart, Fabian Gruss, Alexander E.E. Verkennis, Inge Rondeel, T. van Laar, Pascal Albanese, N.H. Dekker, More Authors Files PDF gkad171.pdf 3.91 MB Close viewer /islandora/object/uuid:9371e760-5c94-4b8c-9912-db2d12963000/datastream/OBJ/view