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Molecular and Cellular Biology, March 2006, p. 1839-1849, Vol. 26, No. 5
0270-7306/06/$08.00+0     doi:10.1128/MCB.26.5.1839-1849.2006
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

Induction of CAF-1 Expression in Response to DNA Strand Breaks in Quiescent Human Cells{dagger}

Arman Nabatiyan, Dávid Szüts,{ddagger} and Torsten Krude*

Department of Zoology, University of Cambridge, Downing Street, Cambridge CB2 3EJ, United Kingdom

Received 24 October 2005/ Returned for modification 20 November 2005/ Accepted 13 December 2005

Genome stability in eukaryotic cells is maintained through efficient DNA damage repair pathways, which have to access and utilize chromatin as their natural template. Here we investigate the role of chromatin assembly factor 1 (CAF-1) and its interacting protein, PCNA, in the response of quiescent human cells to DNA double-strand breaks (DSBs). The expression of CAF-1 and PCNA is dramatically induced in quiescent cells upon the generation of DSBs by the radiomimetic drug bleocin (a bleomycin compound) or by ionizing radiation. This induction depends on DNA-PK. CAF-1 and PCNA are recruited to damaged chromatin undergoing DNA repair of single- and double-strand DNA breaks by the base excision repair and nonhomologous end-joining pathways, respectively, in the absence of extensive DNA synthesis. CAF-1 prepared from repair-proficient quiescent cells after induction by bleocin mediates nucleosome assembly in vitro. Depletion of CAF-1 by RNA interference in bleocin-treated quiescent cells in vivo results in a significant loss of cell viability and an accumulation of DSBs. These results support a novel and essential role for CAF-1 in the response of quiescent human cells to DSBs, possibly by reassembling chromatin following repair of DNA strand breaks.


* Corresponding author. Mailing address: Department of Zoology, University of Cambridge, Downing Street, Cambridge CB2 3EJ, United Kingdom. Phone: 0044 1223 330111. Fax: 0044 1223 336676. E-mail: tk1{at}mole.bio.cam.ac.uk.

{dagger} Supplemental material for this article may be found at http://mcb.asm.org/.

{ddagger} Present address: MRC Laboratory of Molecular Biology, Hills Road, Cambridge CB2 2QH, United Kingdom.


Molecular and Cellular Biology, March 2006, p. 1839-1849, Vol. 26, No. 5
0022-538X/06/$08.00+0     doi:10.1128/MCB.26.5.1839-1849.2006
Copyright © 2006, American Society for Microbiology. All Rights Reserved.




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