ATP-dependent DNA aggregation is a novel function of rat serum albumin.
An ATP-dependent DNA aggregating activity was purified from rat liver by DEAE-cellulose, phosphocellulose, and novobiocin-Sepharose column chromatography. The protein aggregated superhelical, relaxed, single-, or double-stranded DNA in a divalent cation- and ATP-dependent reaction. The DNA aggregating activity was detected by retardation of a DNA-protein complex at the origin on a 1% agarose gel. The protein appeared to exist in solution as a monomer of molecular weight 66,000, and had no DNA polymerase, topoisomerase, recombinase, or ligase activity. The DNA aggregating activity was inhibited by 10 mM nalidixic acid or 1 mM novobiocin but not by 20 mM N-ethylmaleimide or camptothecin. Adenylyl(beta,gamma-methylene)-diphosphonate, adenylyl-imidodiphosphate, or adenosine-5'-O(3-thiotriphosphate) did not substitute for ATP whereas CTP, dTTP, or the ATP analog adenylyl(alpha,beta-methylene)-diphosphonate could replace ATP. The aggregated DNA was only partially dissociated by restriction endonuclease digestion but was completely dissociated by deproteinization with SDS, proteinase K, or chloroform/octanol extraction. On the basis of the molecular weight, thermostability, antigenic property, and amino acid sequence homology in the first 12 positions, we conclude that the rat liver protein is serum albumin and that the ATP-dependent DNA aggregation is a novel function of rat serum albumin.
Nyormoi
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Department of Cell Biology, Baylor College of Medicine, Houston, Texas 77030.
Moses
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D000255:Adenosine Triphosphate / Q000494:pharmacology*; D000818:Animals; D002413:Cations, Divalent; D004247:DNA / Q000187:drug effects / Q000378:metabolism*; D004270:DNA, Circular / Q000187:drug effects / Q000378:metabolism; D004277:DNA, Single-Stranded / Q000187:drug effects / Q000378:metabolism; D004278:DNA, Superhelical / Q000187:drug effects / Q000378:metabolism; D004279:DNA, Viral / Q000187:drug effects / Q000378:metabolism; D004355:Drug Stability; D006358:Hot Temperature; D008099:Liver / Q000737:chemistry*; D046911:Macromolecular Substances; D008970:Molecular Weight; D009268:Nalidixic Acid / Q000494:pharmacology; D009675:Novobiocin / Q000494:pharmacology; D051381:Rats; D011919:Rats, Inbred Strains; D012689:Sequence Homology, Nucleic Acid; D012709:Serum Albumin / Q000737:chemistry / Q000302:isolation & purification / Q000494:pharmacology*
D002413:Cations, Divalent; D004270:DNA, Circular; D004277:DNA, Single-Stranded; D004278:DNA, Superhelical; D004279:DNA, Viral; D046911:Macromolecular Substances; D012709:Serum Albumin; D009675:Novobiocin; D009268:Nalidixic Acid; D000255:Adenosine Triphosphate; D004247:DNA