[Use of benzoyl-naphthyl-DEAE-cellulose chromatography for determining the ability of proteins from Ehrlich ascites carcinoma, binding with single-stranded DNA (SSB-proteins), to destabilize the DNA double helix].
Single-stranded DNA-binding proteins (SSB-proteins) isolated from Ehrlich ascites tumour (EAT) cells were incubated for 30 min at 5 mM NaCl with salmon sperm DNA or [3H]DNA from EAT at the SSB-protein/DNA ratio (w/w) of 0 to 4.5. After addition of sodium dodecyl sulfate up to a 0.05% concentration, the proteins were applied to columns with benzoylated naphthoylated DEAE-cellulose. Double-stranded DNA was eluted by 1 M NaCl; the DNA containing single-stranded regions was eluted by 50% dimethylformamide. There was a progressive lowering of the DNA content in the first eluate and a rise in the second eluate, as could be evidenced from the increase in the SSB-protein/DNA w/w ratio. This effect was more pronounced in the case of homologous DNA and was not coupled with the nuclease activity of SSB proteins. It was concluded that EAT SSB-proteins are "DNA-unwinding" proteins.
Koterov
A N
AN
Andrianova
E P
EP
Filippovich
I V
IV
D000818:Animals; D002286:Carcinoma, Ehrlich Tumor / Q000378:metabolism*; D002853:Chromatography, Liquid / Q000379:methods; D004277:DNA, Single-Stranded / Q000737:chemistry / Q000378:metabolism*; D004268:DNA-Binding Proteins / Q000378:metabolism*; D009363:Neoplasm Proteins / Q000378:metabolism*; D009690:Nucleic Acid Conformation; D012474:Salmon; D014407:Tumor Cells, Cultured
D004277:DNA, Single-Stranded; D004268:DNA-Binding Proteins; D009363:Neoplasm Proteins
Ispol'zovanie khromatografii na benzoil-naftil-DEAE-tselliuloze dlia opredeleniia sposobnosti belkov iz astsitnoĭ kartsinomy Erlikha, sviazyvaiushchikhsia s odnonitevoĭ DNK (SSB-belkov), destabilizirovat'dvoĭnuiu spiral' DNK.