Ku86 preserves chromatin integrity in cells adapted to high NaCl.
Cells adapted to high NaCl have many DNA breaks both in cell culture and in the renal inner medulla in vivo; yet they survive, function, and even proliferate. Here, we show that Ku86 is important for maintaining chromosomal integrity despite the continued presence of DNA breaks. The Ku heterodimer is part of DNA-dependent PK (DNA-PK), a complex that contributes by nonhomologous end joining to repair of double-strand breaks. We demonstrate that cells deficient in Ku86, but not cells deficient in DNA-PKcs (the catalytic subunit of DNA-PK), are hypersensitive to high NaCl as manifested by profound inhibition of proliferation, aberrant mitosis, and increased chromosomal fragmentation. Lower eukaryotes, including the soil nematode Caenorhabditis elegans, lack a DNA-PKcs homologue but are able to adapt to high NaCl. We show that cells of C. elegans adapted to high NaCl have many DNA breaks, similar to the mammalian cells adapted to high NaCl. Ku86 mutant C. elegans as well as C. elegans fed with cku86 dsRNA also display hypersensitivity to high NaCl, characterized by a reduced number of progeny and prolonged generation time in high NaCl. We propose that Ku86 ameliorates the effects of high NaCl-induced DNA breaks in adapted cells by supporting alignment of the broken ends of the DNA and thus maintaining integrity of the fragmented chromatin.
Dmitrieva
Natalia I
NI
Laboratory of Kidney and Electrolyte Metabolism, National Heart, Lung, and Blood Institute, and Experimental Immunology Branch, National Cancer Institute, National Institutes of Health, Bethesda, MD 20892, USA. dmitrien@nhlbi.nih.gov
Celeste
Arkady
A
Nussenzweig
André
A
Burg
Maurice B
MB
D000222:Adaptation, Physiological; D000818:Animals; D034961:Antigens, Nuclear / Q000235:genetics / Q000378:metabolism* / Q000502:physiology; D015153:Blotting, Western; D017173:Caenorhabditis elegans; D002453:Cell Cycle / Q000187:drug effects; D002460:Cell Line; D002843:Chromatin / Q000187:drug effects / Q000235:genetics / Q000502:physiology*; D006224:Cricetinae; D003412:Cricetulus; D020732:Cytogenetic Analysis; D004249:DNA Damage; D004260:DNA Repair / Q000235:genetics / Q000502:physiology*; D051747:DNA-Activated Protein Kinase / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism* / Q000502:physiology; D006801:Humans; D000072200:Ku Autoantigen; D051379:Mice; D009687:Nuclear Proteins / Q000378:metabolism; D034622:RNA Interference; D012098:Reproduction / Q000187:drug effects; D012965:Sodium Chloride / Q000633:toxicity*
D034961:Antigens, Nuclear; D002843:Chromatin; D004268:DNA-Binding Proteins; D009687:Nuclear Proteins; D012965:Sodium Chloride; D051747:DNA-Activated Protein Kinase; C497264:Prkdc protein, mouse; C000606512:Xrcc5 protein, mouse; C000606509:Xrcc6 protein, human; C000606510:Xrcc6 protein, mouse; D000072200:Ku Autoantigen