TY - JOUR
T1 - ATP-dependent DNA aggregation is a novel function of rat serum albumin
AU - Nyormoi, Okot
AU - Moses, Robb E.
N1 - Funding Information:
We thank Drs. Paul Englund and Leonard A. Zwelling for trypanosome Crithidia faxic&ta kinetoplast DNA, Drs. Fred H. Drake and Leroy F. Lui for rabbit antiserum to DNA topoisomerase II, Dr. George Wein-stock for recA protein, and Drs. Fred H. Drake and Leonard A. Zwelling for DNA topoisomerase II enzyme. Supported by USPHS Grant GM24711 and BRSG Grant G13286.
PY - 1991/6
Y1 - 1991/6
N2 - 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(β,γ-methylene)-diphosphonate, adenylyl-imidodiphosphate, or adenosine-5′-O(3-thiotriphosphate) did not substitute for ATP whereas CTP, dTTP, or the ATP analog adenylyl(α,β-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.
AB - 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(β,γ-methylene)-diphosphonate, adenylyl-imidodiphosphate, or adenosine-5′-O(3-thiotriphosphate) did not substitute for ATP whereas CTP, dTTP, or the ATP analog adenylyl(α,β-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.
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U2 - 10.1016/0003-9861(91)90491-Z
DO - 10.1016/0003-9861(91)90491-Z
M3 - Article
C2 - 1898009
AN - SCOPUS:0025849670
VL - 287
SP - 367
EP - 371
JO - Archives of Biochemistry and Biophysics
JF - Archives of Biochemistry and Biophysics
SN - 0003-9861
IS - 2
ER -