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

Clément Rouillon, Bruna V Eckhardt, Leonie Kollenstart, Fabian Gruss, Alexander E E Verkennis, Inge Rondeel, Peter H L Krijger, Giulia Ricci, Alva Biran, Theo van Laar, Charlotte M Delvaux de Fenffe, Georgiana Luppens, Pascal Albanese, Koichi Sato, Richard A Scheltema, Wouter de Laat, Puck Knipscheer, Nynke H Dekker, Anja Groth, Francesca Mattiroli

Research output: Contribution to journal/periodicalArticleScientificpeer-review

12 Citations (Scopus)


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.

Original languageEnglish
Pages (from-to)3770-3792
Number of pages23
JournalNucleic Acids Research
Issue number8
Publication statusPublished - 08 May 2023


  • Histones/metabolism
  • Proliferating Cell Nuclear Antigen/genetics
  • Chromatin Assembly Factor-1/genetics
  • Chromatin/genetics
  • DNA Replication
  • DNA/genetics


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