title: Senescent human fibroblasts have elevated Ku86 proteolytic cleavage activity.
abstract: A proteolytic activity capable of cleaving the Ku86 subunit of Ku protein to two polypeptides, with molecular masses of 69 and 17 kDa in vitro, is present in a human diploid fibroblast (HDF) cell line. The activity is elevated in late-passaged and senescent cells, and the cleaved 69-kDa product seems able to form complex with Ku70 to bind DNA ends. However, further studies indicate that cleavage of Ku86 could be inhibited by including leupeptin in the extraction buffer, and no 69 kDa variant was evident in the cell. In fact, the levels of Ku86, Ku70 and DNA-end binding activity of Ku remained unchanged during replicative senescence. Thus, this study reveals an intriguing protease in HDFs, and also indicates that inconsistent results of Ku86 expression will be obtained if the protease activity is not completely inhibited.
authors.affiliation: Department of Life Science, National Tsin Hua University, Hsinchu, Taiwan.
authors.forename: Y W
authors.initials: YW
authors.lastname: Jeng
authors.forename: H C
authors.initials: HC
authors.lastname: Chao
authors.forename: C F
authors.initials: CF
authors.lastname: Chiu
authors.forename: W G
authors.initials: WG
authors.lastname: Chou
mesh_terms: D034961:Antigens, Nuclear; D002455:Cell Division / Q000502:physiology; D002460:Cell Line; D016922:Cellular Senescence / Q000502:physiology*; D004265:DNA Helicases; D004260:DNA Repair; D004261:DNA Replication; D004268:DNA-Binding Proteins / Q000096:biosynthesis / Q000378:metabolism*; D010450:Endopeptidases / Q000378:metabolism*; D005347:Fibroblasts / Q000166:cytology / Q000201:enzymology* / Q000502:physiology*; D006801:Humans; D006868:Hydrolysis; D066298:In Vitro Techniques; D000072200:Ku Autoantigen; D009687:Nuclear Proteins / Q000096:biosynthesis / Q000378:metabolism*
chemical_list: D034961:Antigens, Nuclear; D004268:DNA-Binding Proteins; D009687:Nuclear Proteins; D010450:Endopeptidases; D004265:DNA Helicases; C106893:XRCC5 protein, human; C000606509:Xrcc6 protein, human; D000072200:Ku Autoantigen