Analysis of DNA breaks, DNA damage response, and apoptosis produced by high NaCl.
We previously reported that, both in cell culture and in the renal inner medulla in vivo, elevating NaCl increased the number of DNA breaks, which persisted as long as NaCl remained high but were rapidly repaired when NaCl was lowered. Furthermore, those breaks did not induce the DNA repair protein gammaH2AX or cause activation of the MRN (Mre11, Rad50, Nbs1) complex. In contrast, others recently reported that high NaCl does induce gammaH2AX and MRN complex formation and concluded that these activities are associated with repair of the DNA (Sheen MR, Kim SW, Jung JY, Ahn JY, Rhee JG, Kwon HM, Woo SK. Am J Physiol Renal Physiol 291: F1014-F1020, 2006). The purpose of the present studies was to resolve the disparity. The important difference is that HeLa cells, which were the main subject of the later report, are much less tolerant of high NaCl than are the mIMCD3 cells, which were our main subject. mIMCD3 cells survive levels of NaCl that kill HeLa cells by apoptosis. Here we demonstrate that in both cell types raising NaCl to a level that the cells survive (higher for mIMCD3 than HeLa) increases DNA breaks without inducing gammaH2AX or activating the MRN complex and that the DNA breaks persist as long as NaCl remains elevated, but are rapidly repaired when it is lowered. Importantly, in both cell types, raising NaCl further to cause apoptosis activates these DNA damage response proteins and greatly fragments DNA, associated with cell death. We conclude that gammaH2AX induction and MRN activation in response to high NaCl are associated with apoptosis, not DNA repair.
Dmitrieva
Natalia I
NI
Laboratory of Kidney and Electrolyte Metabolism, National Heart, Lung and Blood Institute, National Institutes of Health, Dept. of Health and Human Services, 9000 Rockville Pike, Bethesda, MD 20892-1603, USA. dmitrien@nhlbi.nih.gov
Burg
Maurice B
MB
D017209:Apoptosis / Q000187:drug effects*; D020169:Caspases / Q000187:drug effects / Q000378:metabolism; D002455:Cell Division / Q000187:drug effects; D002467:Cell Nucleus / Q000187:drug effects / Q000502:physiology; D003593:Cytoplasm / Q000187:drug effects / Q000502:physiology; D053960:DNA Breaks / Q000187:drug effects*; D004249:DNA Damage / Q000187:drug effects*; D004260:DNA Repair / Q000187:drug effects; D004305:Dose-Response Relationship, Drug; D004361:Drug Tolerance; D004789:Enzyme Activation / Q000187:drug effects; D006367:HeLa Cells / Q000166:cytology / Q000187:drug effects / Q000502:physiology; D006801:Humans; D007679:Kidney Medulla / Q000187:drug effects / Q000503:physiopathology; D012965:Sodium Chloride / Q000494:pharmacology*
D012965:Sodium Chloride; D020169:Caspases