Stimulation of topoisomerase II-mediated DNA damage via a mechanism involving protein thiolation.
The breakage/reunion reaction of DNA topoisomerase II (TOP2) can be interrupted by DNA intercalators (e.g., doxorubicin), enzyme binders (e.g., etoposide), or DNA lesions (e.g., abasic sites) to produce TOP2-mediated DNA damage. Here, we demonstrate that thiol alkylation of TOP2 can also produce TOP2-mediated DNA damage. This conclusion is supported by the following observations using purified TOP2: (1) Thiol-reactive quinones were shown to induce TOP2-mediated DNA cleavage. (2) Thiol-reactive compounds such as N-ethylmaleimide (NEM), disulfiram, and organic disulfides [e.g., 2,2'-dithiobis(5-nitropyridine)] were also shown to induce TOP2-mediated DNA cleavage with similar reaction characteristics as thiol-reactive quinones. (3) TOP2-mediated DNA cleavage induced by thiol-reactive quinones was completely abolished using mutant yeast TOP2 with all cysteine residues replaced with alanine (cysteineless TOP2). These results suggest the possibility that cellular DNA damage could occur indirectly through thiolation of a nuclear protein, TOP2. The implications of this reaction in carcinogenesis and apoptotic cell death are discussed.
Department of Pharmacology, University of Medicine & Dentistry of New Jersey-Robert Wood Johnson Medical School, 675 Hoes Lane, Piscataway, New Jersey 08854-5635, USA.
H
H
Wang
Y
Y
Mao
A Y
AY
Chen
N
N
Zhou
E J
EJ
LaVoie
L F
LF
Liu
D000409:Alanine / Q000235:genetics; D000478:Alkylation; D000818:Animals; D003545:Cysteine / Q000235:genetics; D004249:DNA Damage; D004250:DNA Topoisomerases, Type II / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D004330:Drosophila / Q000201:enzymology; D006801:Humans; D007364:Intercalating Agents / Q000494:pharmacology; D007624:KB Cells; D016297:Mutagenesis, Site-Directed; D009285:Naphthoquinones / Q000494:pharmacology; D011809:Quinones / Q000494:pharmacology; D013438:Sulfhydryl Compounds / Q000378:metabolism* / Q000494:pharmacology; D014407:Tumor Cells, Cultured; D014812:Vitamin K / Q000494:pharmacology
D007364:Intercalating Agents; D009285:Naphthoquinones; D011809:Quinones; D013438:Sulfhydryl Compounds; D014812:Vitamin K; D004250:DNA Topoisomerases, Type II; D003545:Cysteine; D000409:Alanine; C005134:juglone; C014758:plumbagin