DNA damage, signalling and repair after exposure of cells to the sulphur mustard analogue 2-chloroethyl ethyl sulphide.
Sulphur mustard (SM) is a blistering agent that is directly toxic to the skin and mucosal surfaces of the eye and respiratory system. Symptoms take several hours to develop and the mechanism of action is poorly understood although SM is able to alkylate nucleic acids and proteins. The ability of SM to form adducts with DNA has been documented, although there are limited data demonstrating how cells respond to this insult to repair the damage. This study used the sulphur mustard surrogate 2-chloroethyl ethyl sulphide (CEES) to identify DNA damage repair pathways and signalling events that are activated after exposure to the agent. A dose-dependent increase in DNA damage was observed in TK6 lymphoblastoid cells, which was associated with a loss of cell viability. Using both model human lymphoblastoid cell lines and pharmacological inhibitors, it was found that DNA damage induced by CEES was repaired by base excision repair (BER) and nucleotide excision repair (NER) pathways. Finally, CEES was found to induce the phosphorylation of p53 and Chk2 and these events were mediated by both the ATM ataxia telangiectasia mutated and ATR (ATM and Rad-3 related) protein kinases.
Jowsey
Paul A
PA
Medical Toxicology Centre, Wolfson Unit, Newcastle University, Claremont Place, Newcastle upon Tyne NE2 4AA, United Kingdom. p.a.jowsey@ncl.ac.uk
Williams
Faith M
FM
Blain
Peter G
PG
D064007:Ataxia Telangiectasia Mutated Proteins; D015153:Blotting, Western; D018797:Cell Cycle Proteins / Q000235:genetics; D045744:Cell Line, Tumor; D002470:Cell Survival / Q000187:drug effects; D064447:Checkpoint Kinase 2; D020552:Comet Assay; D004247:DNA / Q000378:metabolism; D004249:DNA Damage; D004260:DNA Repair / Q000187:drug effects*; D004268:DNA-Binding Proteins / Q000235:genetics; D006801:Humans; D009097:Multienzyme Complexes / Q000235:genetics; D009151:Mustard Gas / Q000031:analogs & derivatives* / Q000633:toxicity; D043264:Phosphodiesterase I / Q000235:genetics; D010727:Phosphoric Diester Hydrolases; D010766:Phosphorylation; D017346:Protein Serine-Threonine Kinases / Q000187:drug effects / Q000235:genetics / Q000502:physiology; D011755:Pyrophosphatases / Q000235:genetics; D015398:Signal Transduction / Q000187:drug effects*; D016159:Tumor Suppressor Protein p53 / Q000187:drug effects / Q000235:genetics / Q000502:physiology; D025521:Tumor Suppressor Proteins / Q000235:genetics; D000099189:Lysophospholipase D
D064007:Ataxia Telangiectasia Mutated Proteins; D018797:Cell Cycle Proteins; D064447:Checkpoint Kinase 2; D004247:DNA; D004268:DNA-Binding Proteins; D009097:Multienzyme Complexes; D009151:Mustard Gas; D043264:Phosphodiesterase I; D010727:Phosphoric Diester Hydrolases; D017346:Protein Serine-Threonine Kinases; D011755:Pyrophosphatases; D016159:Tumor Suppressor Protein p53; D025521:Tumor Suppressor Proteins; D000099189:Lysophospholipase D; C031278:2-chloroethyl ethyl sulfide; C576324:ATM protein, human; C578634:CHEK2 protein, human