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Molecular and Cellular Biology, February 2000, p. 1206-1218, Vol. 20, No. 4
0270-7306/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

A CAF-1-PCNA-Mediated Chromatin Assembly Pathway Triggered by Sensing DNA Damage

Jonathan G. Moggs,1 Paola Grandi,1 Jean-Pierre Quivy,1 Zophonías O. Jónsson,2 Ulrich Hübscher,2 Peter B. Becker,3 and Geneviève Almouzni1,*

Institut Curie/Section de Recherche UMR 218 du CNRS, 75231 Paris cedex 05, France1; Institute of Veterinary Biochemistry, University of Zürich-Irchel, CH-8057 Zurich, Switzerland2; and European Molecular Biology Laboratory, 69012 Heidelberg, Germany3

Received 8 June 1999/Returned for modification 10 September 1999/Accepted 19 November 1999

Sensing DNA damage is crucial for the maintenance of genomic integrity and cell cycle progression. The participation of chromatin in these events is becoming of increasing interest. We show that the presence of single-strand breaks and gaps, formed either directly or during DNA damage processing, can trigger the propagation of nucleosomal arrays. This nucleosome assembly pathway involves the histone chaperone chromatin assembly factor 1 (CAF-1). The largest subunit (p150) of this factor interacts directly with proliferating cell nuclear antigen (PCNA), and critical regions for this interaction on both proteins have been mapped. To isolate proteins specifically recruited during DNA repair, damaged DNA linked to magnetic beads was used. The binding of both PCNA and CAF-1 to this damaged DNA was dependent on the number of DNA lesions and required ATP. Chromatin assembly linked to the repair of single-strand breaks was disrupted by depletion of PCNA from a cell-free system. This defect was rescued by complementation with recombinant PCNA, arguing for role of PCNA in mediating chromatin assembly linked to DNA repair. We discuss the importance of the PCNA-CAF-1 interaction in the context of DNA damage processing and checkpoint control.

* Corresponding author. Mailing address: Institut Curie/Section de Recherche UMR 218 du CNRS, 26 rue d'Ulm, 75231 Paris cedex 05, France. Phone: 00 33 1 42 34 64 10. Fax: 00 33 1 42 34 64 21. E-mail: almouzni{at}curie.fr.

Molecular and Cellular Biology, February 2000, p. 1206-1218, Vol. 20, No. 4
0270-7306/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


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