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Molecular and Cellular Biology, February 2000, p. 1206-1218, Vol. 20, No. 4
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.
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
*
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.
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