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Mutagenesis vol. 18 no. 5 pp. 411-416, September 2003
© 2003 UK Environmental Mutagen Society/Oxford University Press

Local DNA damage by proton microbeam irradiation induces poly(ADP-ribose) synthesis in mammalian cells

Laurence Tartier, Catherine Spenlehauer, Heidi C. Newman1, Melvyn Folkard1, Kevin M. Prise1, Barry D. Michael1, Josiane Ménissier-de Murcia and Gilbert de Murcia2

Unité 9003 du CNRS, Laboratoire Conventionné avec le Commissariat à l’Energie Atomique, Ecole Supérieure de Biotechnologie de Strasbourg, Boulevard Sebastien Brant, BP 10413, F-67412 Illkirch-Graffenstaden, France and 1Gray Cancer Institute, PO Box 100, Mount Vernon Hospital, Northwood, Middlesex HA6 2JR, UK

Cellular recovery from ionizing radiation (IR)-induced damage involves poly(ADP-ribose) polymerase (PARP-1 and PARP-2) activity, resulting in the induction of a signalling network responsible for the maintenance of genomic integrity. In the present work, a charged particle microbeam delivering 3.2 MeV protons from a Van de Graaff accelerator has been used to locally irradiate mammalian cells. We show the immediate response of PARPs to local irradiation, concomitant with the recruitment of ATM and Rad51 at sites of DNA damage, both proteins being involved in DNA strand break repair. We found a co-localization but no connection between two DNA damage-dependent post-translational modifications, namely poly(ADP-ribosyl)ation of nuclear proteins and phosphorylation of histone H2AX. Both of them, however, should be considered and used as bona fide immediate sensitive markers of IR damage in living cells. This technique thus provides a powerful approach aimed at understanding the interactions between the signals originating from sites of DNA damage and the subsequent activation of DNA strand break repair mechanisms

2To whom correspondence should be addressed. Tel: +33 3 9024 4707; Fax: +33 3 9024 4686; Email: demurcia{at}esbs.u-strasbg.fr

Received on October 31, 2002 revised and accepted on May 19, 2003


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