TY - JOUR

T1 - CAF-1 deposits newly synthesized histones during DNA replication using distinct mechanisms on the leading and lagging strands

AU - Rouillon, Clément

AU - Eckhardt, Bruna V

AU - Kollenstart, Leonie

AU - Gruss, Fabian

AU - Verkennis, Alexander E E

AU - Rondeel, Inge

AU - Krijger, Peter H L

AU - Ricci, Giulia

AU - Biran, Alva

AU - van Laar, Theo

AU - Delvaux de Fenffe, Charlotte M

AU - Luppens, Georgiana

AU - Albanese, Pascal

AU - Sato, Koichi

AU - Scheltema, Richard A

AU - de Laat, Wouter

AU - Knipscheer, Puck

AU - Dekker, Nynke H

AU - Groth, Anja

AU - Mattiroli, Francesca

N1 - © The Author(s) 2023. Published by Oxford University Press on behalf of Nucleic Acids Research.

PY - 2023

Y1 - 2023

N2 - 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.

AB - 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.

U2 - 10.1093/nar/gkad171

DO - 10.1093/nar/gkad171

M3 - Journal article

C2 - 36942484

VL - 51

SP - 3770

EP - 3792

JO - Nucleic Acids Research

JF - Nucleic Acids Research

SN - 0305-1048

IS - 8

ER -