Characterization of a DNA damage-recognition protein mammalian cells that binds specifically to intrastrand d(GpG) and d(ApG) DNA adducts of the anticancer drug cisplatin

Journal Article · Tue Jun 19 04:00:00 EDT 1990 · Biochemistry; (USA)

DOI:https://doi.org/10.1021/bi00476a032· OSTI ID:6287133

Donahue, B A; Augot, M; Bellon, S F; Treiber, D K; Toney, J H; Lippard, S J; Essigmann, J M ^[1]

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1. Massachusetts Institute of Technology, Cambridge (USA)

A factor has been identified in extracts from human HeLa and hamster V79 cells that retards the electrophoretic mobility of several DNA restriction fragments modified with the antitumor drug cis-diamminedichloroplatinum(II) (cisplatin). Binding of the factor to cisplatin-modified DNA was sensitive to pretreatment with proteinase K, establishing that the factor is a protein. Gel mobility shifts were observed with probes containing as few as seven Pt atoms per kilobase of duplex DNA. By competition experiments the dissociation constant, K{sub d}, of the protein from cisplatin-modified DNA was estimated to be (1-20) {times} 10{sup {minus}10} M. Protein binding is selective for DNA modified with cisplatin, (Pt(en)Cl{sub 2}) (en, ethylenediamine), and (Pt(dach)Cl{sub 2}) (dach, 1,2-diaminocyclohexane) but not with chemotherapeutically inactive trans-diamminedichloroplatinum(II) or monofunctionally coordinating (Pt(dien)Cl)Cl (dien, diethylenetriamine) complexes. The protein binds specifically to 1,2-intrastrand d(GpG) and d(ApG) cross-links formed by cisplatin. The apparent molecular weight of the protein is 91,000, as determined by sucrose gradient centrifugation of a preparation partially purified by ammonium sulfate fractionation. Binding of the protein to platinum-modified DNA does not require cofactors but is sensitive to treatment with 5 mM MnCl{sub 2}, CdCl{sub 2}, CoCl{sub 2}, or ZnCl{sub 2} and with 1 mM HgCl{sub 2}. This protein, alone or in conjunction with other cellular constituents, could be of general importance in the initial stages of processing of mammalian DNA damaged by cisplatin or other genotoxic agents and may belong to a wider class of such cellular damage-recognition proteins (DRPs).

• OSTI ID: 6287133
• Journal Information: Biochemistry; (USA), Journal Name: Biochemistry; (USA) Vol. 29:24; ISSN 0006-2960; ISSN BICHA
• Country of Publication: United States
• Language: English

Related Subjects

560120 -- Radiation Effects on Biochemicals
Cells
& Tissue Culture
560300* -- Chemicals Metabolism & Toxicology
63 RADIATION, THERMAL, AND OTHER ENVIRON. POLLUTANT EFFECTS ON LIVING ORGS. AND BIOL. MAT.
ADDUCTS
ANIMAL CELLS
ANIMALS
BIOLOGICAL EFFECTS
CADMIUM CHLORIDES
CADMIUM COMPOUNDS
CADMIUM HALIDES
CHEMICAL REACTIONS
CHLORIDES
CHLORINE COMPOUNDS
COBALT CHLORIDES
COBALT COMPOUNDS
CONNECTIVE TISSUE CELLS
CROSS-LINKING
DNA ADDUCTS
ELECTROMAGNETIC RADIATION
ELECTROPHORESIS
FIBROBLASTS
HALIDES
HALOGEN COMPOUNDS
HAMSTERS
HELA CELLS
MAMMALS
MANGANESE CHLORIDES
MANGANESE COMPOUNDS
MANGANESE HALIDES
NUCLEIC ACIDS
OLIGONUCLEOTIDES
ORGANIC COMPOUNDS
PLATINUM COMPOUNDS
POLYMERIZATION
PROTEINS
RADIATIONS
RODENTS
SEDIMENTATION
SOMATIC CELLS
TRANSITION ELEMENT COMPOUNDS
ULTRAVIOLET RADIATION
VERTEBRATES
ZINC CHLORIDES
ZINC COMPOUNDS
ZINC HALIDES