Characterization of a duocarmycin-DNA adduct-recognizing protein in cancer cells.
Duocarmycins have been reported to derive their potent antitumor activity through a sequence-selective minor groove alkylation of N3 adenine in double-stranded DNA. We have used gel mobility shift assays to detect proteins that bind to DNA treated in vitro with duocarmycin SA and identified a protein, named duocarmycin-DNA adduct recognizing protein (DARP), which binds with increased affinity to duocarmycin-damaged DNA. Examination with partially purified DARP revealed that the protein recognized not only the DNA adduct of structurally related drug, CC-1065, but unexpectedly, the protein also recognized the DNA adduct of another chemotype of minor groove binder, anthramycin. These results demonstrate that DARP recognizes the structural alteration of DNA induced by these potent DNA-alkylating drugs, suggesting the possibility that the protein might modulate the antitumor activity of these drugs.
Asai
A
A
Tokyo Research Laboratories, Kyowa Hakko Kogyo Co., Ltd., Japan. aasai@kyowa.co.jp
Yano
K
K
Mizukami
T
T
Nakano
H
H
D000818:Animals; D000876:Anthramycin / Q000378:metabolism; D000903:Antibiotics, Antineoplastic / Q000378:metabolism*; D017209:Apoptosis; D001667:Binding, Competitive; D002417:Cattle; D002467:Cell Nucleus / Q000737:chemistry / Q000378:metabolism; D004247:DNA / Q000378:metabolism; D018736:DNA Adducts / Q000737:chemistry / Q000378:metabolism*; D004268:DNA-Binding Proteins / Q000737:chemistry* / Q000302:isolation & purification; D000080890:Duocarmycins; D004591:Electrophoresis, Polyacrylamide Gel; D006367:HeLa Cells; D006801:Humans; D007211:Indoles; D007933:Leucomycins / Q000378:metabolism; D009687:Nuclear Proteins / Q000737:chemistry* / Q000302:isolation & purification; D009841:Oligonucleotides / Q000378:metabolism; D011758:Pyrroles / Q000378:metabolism; D012261:Ribonucleoproteins; D013950:Thymus Gland / Q000378:metabolism; D014407:Tumor Cells, Cultured; D034664:Heterogeneous-Nuclear Ribonucleoprotein L
D000876:Anthramycin; D000903:Antibiotics, Antineoplastic; D004247:DNA; D018736:DNA Adducts; D004268:DNA-Binding Proteins; D000080890:Duocarmycins; D007211:Indoles; D007933:Leucomycins; D009687:Nuclear Proteins; D009841:Oligonucleotides; D011758:Pyrroles; D012261:Ribonucleoproteins; C400563:HNRNPL protein, human; D034664:Heterogeneous-Nuclear Ribonucleoprotein L; C066098:duocarmycin SA; C018782:CC 1065