An inducible Ku86-degrading serine protease in human cells.
The Ku autoantigen has been implicated in a number of cellular functions including growth control, immunoglobulin gene rearrangement and DNA repair. A variant truncated form of Ku86, with an apparent molecular weight of 70 kDa, has been reported to be present in many human cell types. We have previously shown that the amount of variant Ku86 is strongly increased in human peripheral blood mononuclear cells (PBMC) by storage of blood prior to isolation of the PBMC. In this study we report that formation of variant Ku86 in protein extracts is mediated by an inducible trypsin-like serine protease with a higher concentration in the nuclear compartment, as compared with the cytoplasm. However, experiments with SDS-PAGE assay of whole cells yielded no evidence of truncated Ku86, suggesting that the protease is not active in intact cells, but is exerting a marked activity during the protein extraction procedure. Interestingly, the protease level became markedly reduced upon transfer of the cells to growth medium. Protease induction did not correlate with apoptosis, necrotic cell death or with signs of general proteolysis or cytotoxicity. Our findings have methodological implications for the interpretation of experimental Ku86 data, and suggest that this protease may play a role for cellular regulation of Ku function.
Sallmyr
Annahita
A
Department of Medical Microbiology, Lund University, Malmö University Hospital, 205 02 Malmö, Sweden.
Du
Liping
L
Bredberg
Anders
A
D034961:Antigens, Nuclear / Q000378:metabolism*; D002467:Cell Nucleus / Q000201:enzymology; D004265:DNA Helicases; D004268:DNA-Binding Proteins / Q000378:metabolism*; D004591:Electrophoresis, Polyacrylamide Gel; D004790:Enzyme Induction; D006801:Humans; D000072200:Ku Autoantigen; D007963:Leukocytes, Mononuclear / Q000201:enzymology*; D012697:Serine Endopeptidases / Q000096:biosynthesis / Q000378:metabolism*; D015842:Serine Proteinase Inhibitors / Q000378:metabolism
D034961:Antigens, Nuclear; D004268:DNA-Binding Proteins; D015842:Serine Proteinase Inhibitors; D012697:Serine Endopeptidases; D004265:DNA Helicases; C106893:XRCC5 protein, human; C000606509:Xrcc6 protein, human; D000072200:Ku Autoantigen