Local DNA damage by proton microbeam irradiation induces poly(ADP-ribose) synthesis in mammalian cells.
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
Tartier
Laurence
L
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.
Spenlehauer
Catherine
C
Newman
Heidi C
HC
Folkard
Melvyn
M
Prise
Kevin M
KM
Michael
Barry D
BD
Ménissier-de Murcia
Josiane
J
de Murcia
Gilbert
G
D000818:Animals; D064007:Ataxia Telangiectasia Mutated Proteins; D018797:Cell Cycle Proteins; D002478:Cells, Cultured; D004249:DNA Damage; D004260:DNA Repair / Q000502:physiology; D004268:DNA-Binding Proteins / Q000378:metabolism / Q000528:radiation effects; D004307:Dose-Response Relationship, Radiation; D006367:HeLa Cells / Q000528:radiation effects; D006657:Histones / Q000378:metabolism / Q000528:radiation effects; D006801:Humans; D008322:Mammals; D009154:Mutation; D010766:Phosphorylation; D011064:Poly Adenosine Diphosphate Ribose / Q000096:biosynthesis*; D011065:Poly(ADP-ribose) Polymerases / Q000235:genetics / Q000378:metabolism / Q000528:radiation effects; D011499:Protein Processing, Post-Translational; D017346:Protein Serine-Threonine Kinases / Q000378:metabolism / Q000528:radiation effects; D011522:Protons; D051135:Rad51 Recombinase; D011839:Radiation, Ionizing; D025521:Tumor Suppressor Proteins
D018797:Cell Cycle Proteins; D004268:DNA-Binding Proteins; D006657:Histones; D011522:Protons; D025521:Tumor Suppressor Proteins; D011064:Poly Adenosine Diphosphate Ribose; C483041:PARP2 protein, human; D011065:Poly(ADP-ribose) Polymerases; C576324:ATM protein, human; D064007:Ataxia Telangiectasia Mutated Proteins; D017346:Protein Serine-Threonine Kinases; C495153:RAD51 protein, human; D051135:Rad51 Recombinase