title: High NaCl causes Mre11 to leave the nucleus, disrupting DNA damage signaling and repair.
abstract: High NaCl causes DNA double-strand breaks and cell cycle arrest, but the mechanism of its genotoxicity has been unclear. In this study, we describe a novel mechanism that contributes to this genotoxicity. The Mre11 exonuclease complex is a central component of DNA damage response. This complex assembles at sites of DNA damage, where it processes DNA ends for subsequent activation of repair and initiates cell cycle checkpoints. However, this does not occur with DNA damage caused by high NaCl. Rather, following high NaCl, Mre11 exits from the nucleus, DNA double-strand breaks accumulate in the S and G2 phases of the cell cycle, and DNA repair is inhibited. Furthermore, the exclusion of Mre11 from the nucleus by high NaCl persists following UV or ionizing radiation, also preventing DNA repair in response to those stresses, as evidenced by absence of H2AX phosphorylation at places of DNA damage and by impaired repair of damaged reporter plasmids. Activation of chk1 by phosphorylation on Ser345 generally is required for DNA damage-induced cell cycle arrest. However, chk1 does not become phosphorylated during high NaCl-induced cell cycle arrest. Also, high NaCl prevents ionizing and UV radiation-induced phosphorylation of chk1, but cell cycle arrest still occurs, indicating the existence of alternative mechanisms for the S and G2/M delays. DNA breaks that occur normally during processes such as DNA replication and transcription, as well as damages to DNA induced by genotoxic stresses, ordinarily are rapidly repaired. We propose that inhibition of this repair by high NaCl results in accumulation of DNA damage, accounting for the genotoxicity of high NaCl, and that cell cycle delay induced by high NaCl slows accumulation of DNA damage until the DNA damage-response network can be reactivated.
authors.affiliation: Laboratory of Kidney and Electrolyte Metabolism, National Heart, Lung, and Blood Institute, National Institutes of Health, Bldg. 10, Rm. 6N260, Bethesda, MD 20892, USA. dmitrien@nhlbi.nih.gov
authors.forename: Natalia I
authors.initials: NI
authors.lastname: Dmitrieva
authors.forename: Dmitry V
authors.initials: DV
authors.lastname: Bulavin
authors.forename: Maurice B
authors.initials: MB
authors.lastname: Burg
mesh_terms: D000818:Animals; D002467:Cell Nucleus / Q000378:metabolism; D002478:Cells, Cultured; D000071877:Checkpoint Kinase 1; D004249:DNA Damage / Q000187:drug effects / Q000502:physiology*; D004260:DNA Repair / Q000187:drug effects / Q000502:physiology*; D045643:DNA Repair Enzymes; D004268:DNA-Binding Proteins / Q000378:metabolism*; D016195:G2 Phase / Q000187:drug effects / Q000235:genetics; D006657:Histones / Q000378:metabolism; D007685:Kidney Tubules, Collecting / Q000166:cytology / Q000502:physiology*; D000076228:MRE11 Homologue Protein; D051379:Mice; D008938:Mitosis / Q000187:drug effects / Q000235:genetics; D010766:Phosphorylation; D011494:Protein Kinases / Q000378:metabolism; D011839:Radiation, Ionizing; D016196:S Phase / Q000187:drug effects / Q000235:genetics; D012965:Sodium Chloride / Q000494:pharmacology*; D014466:Ultraviolet Rays
chemical_list: D000071877:Checkpoint Kinase 1; D045643:DNA Repair Enzymes; D004268:DNA-Binding Proteins; D006657:Histones; D000076228:MRE11 Homologue Protein; D011494:Protein Kinases; D012965:Sodium Chloride; C512689:Mre11a protein, mouse; C000605869:Chek1 protein, mouse; C475961:gamma-H2AX protein, mouse