The RING-H2-finger protein APC11 as a target of hydrogen peroxide.
The anaphase-promoting complex (APC) is a ubiquitin-protein ligase (E3) that targets cell cycle regulators such as cyclin B and securin for degradation. The APC11 subunit functions as the catalytic core of this complex and mediates the transfer of ubiquitin from a ubiquitin-conjugating enzyme (E2) to the substrate. APC11 contains a RING-H2-finger domain, which includes one histidine and seven cysteine residues that coordinate two Zn(2+) ions. We now show that exposure of purified APC11 to H(2)O(2) (0.1 to 1 mM) induced the release of bound zinc as a result of the oxidation of cysteine residues. It also impaired the physical interaction between APC11 and the E2 enzyme Ubc4 as well as inhibited the ubiquitination of cyclin B1 by APC11. The release of HeLa cells from metaphase arrest in the presence of exogenous H(2)O(2) inhibited the ubiquitination of cyclin B1 as well as the degradation of cyclin B1 and securin that were apparent in the absence of H(2)O(2). The presence of H(2)O(2) also blocked the co-immunoprecipitation of Ubc4 with APC11 and delayed the exit of cells from mitosis. Inhibition of APC11 function by H(2)O(2) thus likely contributes to the delay in cell cycle progression through mitosis that is characteristic of cells subjected to oxidative stress.
Chang
Tong-Shin
TS
Laboratory of Cell Signaling, National Heart, Lung, and Blood Institute, National Institutes of Health, Bethesda, MD 20892, USA.
Jeong
Woojin
W
Lee
Duck-Yeon
DY
Cho
Chun-Seok
CS
Rhee
Sue Goo
SG
D000705:Anaphase; D064173:Anaphase-Promoting Complex-Cyclosome; D000818:Animals; D064198:Apc11 Subunit, Anaphase-Promoting Complex-Cyclosome; D002384:Catalysis; D020134:Catalytic Domain; D018929:Cell Culture Techniques; D002453:Cell Cycle; D018797:Cell Cycle Proteins; D019926:Cyclin B / Q000378:metabolism; D056744:Cyclin B1; D003545:Cysteine / Q000737:chemistry; D004926:Escherichia coli / Q000378:metabolism; D005434:Flow Cytometry; D005609:Free Radicals; D006367:HeLa Cells; D006639:Histidine / Q000737:chemistry; D006801:Humans; D006861:Hydrogen Peroxide / Q000737:chemistry / Q000378:metabolism* / Q000494:pharmacology; D047468:Immunoprecipitation; D007477:Ions; D051379:Mice; D015877:Miosis; D009363:Neoplasm Proteins / Q000378:metabolism / Q000502:physiology*; D009687:Nuclear Proteins / Q000378:metabolism / Q000502:physiology*; D018384:Oxidative Stress; D010100:Oxygen / Q000378:metabolism; D010957:Plasmids / Q000378:metabolism; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D011994:Recombinant Proteins / Q000737:chemistry; D064249:Securin; D013438:Sulfhydryl Compounds / Q000737:chemistry; D013997:Time Factors; D014162:Transfection; D044763:Ubiquitin-Conjugating Enzymes / Q000378:metabolism; D043743:Ubiquitin-Protein Ligase Complexes; D015032:Zinc / Q000737:chemistry
C534699:CCNB1 protein, human; C534700:Ccnb1 protein, mouse; D018797:Cell Cycle Proteins; D019926:Cyclin B; D056744:Cyclin B1; D005609:Free Radicals; D007477:Ions; D009363:Neoplasm Proteins; D009687:Nuclear Proteins; D011994:Recombinant Proteins; D064249:Securin; D013438:Sulfhydryl Compounds; C578002:pituitary tumor-transforming protein 1, human; D006639:Histidine; D006861:Hydrogen Peroxide; D044763:Ubiquitin-Conjugating Enzymes; D043743:Ubiquitin-Protein Ligase Complexes; C577351:ANAPC11 protein, human; D064173:Anaphase-Promoting Complex-Cyclosome; D064198:Apc11 Subunit, Anaphase-Promoting Complex-Cyclosome; D015032:Zinc; D003545:Cysteine; D010100:Oxygen