Isolation of cDNAs encoding a human protein that binds selectively to DNA modified by the anticancer drug cis-diammine-dichloroplatinum(II)

Journal Article · Tue Oct 31 23:00:00 EST 1989 · Proceedings of the National Academy of Sciences of the United States of America; (USA)

DOI:https://doi.org/10.1073/pnas.86.21.8328· OSTI ID:7165669

Toney, J H; Kellett, P J; Bruhn, S L; Essigmann, J M; Lippard, S J; Dohahue, B A ^[1]

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1. Massachusetts Institute of Technology, Cambridge (USA)

DNA modified by the antitumor drug cis-diamminedichloroplatinum(II) (cis-DDP or cisplatin) was used to identify a factor in mammalian cells that binds to cis-DDP-damaged DNA and hence may play a role in repair. This factor selectivity recognizes double-stranded DNA fragments modified by cis-DDP or (Pt(en)Cl{sub 2}) (en, ethylenediamine). Little or no binding occurs to unmodified double-stranded DNA or to DNA modified with the clinically ineffective compounds trans-DDP and (Pt(dien)Cl)Cl (dien, diethylenetriamine). Low levels of binding to single-stranded DNA modified by cis-DDP are observed. The apparent molecular mass of the factor in a variety of mammalian cells is {approx} 100 kDa, as determined by modified Western blotting. Two recombinant phage have been isolated from a human B-cell {lambda}gt11 library by using a cis-DDP-modified DNA restriction fragments as a probe. The two clones have insert sizes of 1.88 and 1.44 kilobases and are aligned at their 5{prime} ends. The polypeptides encoded by the recombinant phage exhibit DNA binding properties similar to those of the cellular factor identified in crude extracts prepared from mammalian cells. Northern analysis with one of the clones revealed an mRNA of 2.8 kilobases that is conserved in humans and rodents. The methods used here should be applicable in studies of other damage-specific DNA binding proteins.

• OSTI ID: 7165669
• Journal Information: Proceedings of the National Academy of Sciences of the United States of America; (USA), Journal Name: Proceedings of the National Academy of Sciences of the United States of America; (USA) Vol. 86:21; ISSN 0027-8424; ISSN PNASA
• Country of Publication: United States
• Language: English

Related Subjects

550201* -- Biochemistry-- Tracer Techniques
59 BASIC BIOLOGICAL SCIENCES
BACTERIOPHAGES
BETA DECAY RADIOISOTOPES
BETA-MINUS DECAY RADIOISOTOPES
BIOLOGICAL RECOVERY
BIOLOGICAL REPAIR
CHEMICAL REACTIONS
CROSS-LINKING
DAYS LIVING RADIOISOTOPES
DNA
DNA HYBRIDIZATION
DNA REPAIR
GENETIC MAPPING
HELA CELLS
HYBRIDIZATION
ISOTOPES
LIGHT NUCLEI
MAPPING
MESSENGER-RNA
MICROORGANISMS
MOLECULAR BIOLOGY
NUCLEI
NUCLEIC ACIDS
ODD-ODD NUCLEI
ORGANIC COMPOUNDS
PARASITES
PEPTIDES
PHOSPHORUS 32
PHOSPHORUS ISOTOPES
PLATINUM COMPOUNDS
POLYMERIZATION
POLYPEPTIDES
PROTEINS
RADIOISOTOPES
RECOMBINANT DNA
RECOVERY
REPAIR
RNA
TRANSITION ELEMENT COMPOUNDS
VIRUSES