Proteasomal degradation of oxidatively damaged endogenous histones in K562 human leukemic cells.
A number of antitumor drugs act via the oxidation of nuclear material in the tumor cell. It is therefore important to know if tumor cells can effectively and precisely cope not only with oxidatively induced DNA damage, but also with nuclear protein oxidation. In this study, we investigated the endogenous degradation of oxidatively damaged histones in K562 human leukemic cells after oxidative challenge and demonstrated a link to the overall cellular stress response pathways by poly-ADP-ribose-polymerase (PARP). After an oxidative challenge, endogenous nuclear protein degradation, as well as histone degradation, was enhanced. Among the histone fractions, histone H1 revealed the highest degradation rate, and more than 85% of the total degraded H1 disappeared in the first 30 min after oxidative challenge. Short-term degradation of histones up to 30 min, as well as long-term degradation up to 48 h after oxidative challenge, was significantly reduced in the presence of the PARP inhibitor 3-aminobenzamide, and nearly completely abrogated by the selective proteasome inhibitor lactacystin. Immunoprecipitation experiments indicated that the proteasome specifically degraded oxidized histones. Thus, we show that the nuclear proteosome system in tumor cells is capable of preventing the accumulation of oxidized proteins in this compartment and may suggest further treatment strategies to effectively interfere with the protein "repair" and replacement strategies of tumor cells.
Ullrich
O
O
Dept. of Cell- and Neurobiology, Institute of Anatomy, Humboldt University Berlin, Schumannstrasse 20/21, D-10098 Berlin, Germany.
Grune
T
T
D000111:Acetylcysteine / Q000031:analogs & derivatives* / Q000378:metabolism / Q000494:pharmacology*; D001549:Benzamides / Q000378:metabolism / Q000494:pharmacology; D002467:Cell Nucleus / Q000378:metabolism / Q000648:ultrastructure; D003546:Cysteine Endopeptidases / Q000378:metabolism* / Q000648:ultrastructure; D004791:Enzyme Inhibitors / Q000378:metabolism / Q000494:pharmacology; D006657:Histones / Q000378:metabolism*; D006801:Humans; D020014:K562 Cells; D015464:Leukemia, Myelogenous, Chronic, BCR-ABL Positive / Q000378:metabolism*; D009097:Multienzyme Complexes / Q000037:antagonists & inhibitors* / Q000378:metabolism* / Q000648:ultrastructure; D010084:Oxidation-Reduction; D018384:Oxidative Stress; D011064:Poly Adenosine Diphosphate Ribose / Q000378:metabolism; D000067856:Poly(ADP-ribose) Polymerase Inhibitors; D011065:Poly(ADP-ribose) Polymerases / Q000378:metabolism*; D046988:Proteasome Endopeptidase Complex
D001549:Benzamides; D004791:Enzyme Inhibitors; D006657:Histones; D009097:Multienzyme Complexes; D000067856:Poly(ADP-ribose) Polymerase Inhibitors; C067713:lactacystin; D011064:Poly Adenosine Diphosphate Ribose; C025160:3-aminobenzamide; D011065:Poly(ADP-ribose) Polymerases; D003546:Cysteine Endopeptidases; D046988:Proteasome Endopeptidase Complex; D000111:Acetylcysteine