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

Laurence Tartier; Catherine Spenlehauer; Heidi C Newman; Melvyn Folkard; Kevin M Prise; Barry D Michael; Josiane Ménissier-de Murcia; Gilbert de Murcia

doi:10.1093/mutage/geg015

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

Mutagenesis. 2003 Sep;18(5):411-6. doi: 10.1093/mutage/geg015.

Authors

Laurence Tartier¹, Catherine Spenlehauer, Heidi C Newman, Melvyn Folkard, Kevin M Prise, Barry D Michael, Josiane Ménissier-de Murcia, Gilbert de Murcia

Affiliation

¹ 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.

PMID: 12960408
DOI: 10.1093/mutage/geg015

Abstract

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

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Ataxia Telangiectasia Mutated Proteins
Cell Cycle Proteins
Cells, Cultured
DNA Damage*
DNA Repair / physiology
DNA-Binding Proteins / metabolism
DNA-Binding Proteins / radiation effects
Dose-Response Relationship, Radiation
HeLa Cells / radiation effects
Histones / metabolism
Histones / radiation effects
Humans
Mammals
Mutation
Phosphorylation
Poly Adenosine Diphosphate Ribose / biosynthesis*
Poly(ADP-ribose) Polymerases / genetics
Poly(ADP-ribose) Polymerases / metabolism
Poly(ADP-ribose) Polymerases / radiation effects
Protein Processing, Post-Translational
Protein Serine-Threonine Kinases / metabolism
Protein Serine-Threonine Kinases / radiation effects
Protons
Rad51 Recombinase
Radiation, Ionizing*
Tumor Suppressor Proteins

Substances

Cell Cycle Proteins
DNA-Binding Proteins
Histones
Protons
Tumor Suppressor Proteins
Poly Adenosine Diphosphate Ribose
PARP2 protein, human
Poly(ADP-ribose) Polymerases
ATM protein, human
Ataxia Telangiectasia Mutated Proteins
Protein Serine-Threonine Kinases
RAD51 protein, human
Rad51 Recombinase