Isolation and Characterization of DNA Polymerases from Mature Oocytes of the Starfish, Asterina pectinifera
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概要
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Four distinctive DNA polymerases have been isolated from mature oocytes of the starfish, Asterina pectinifera, and characterized. The N-ethylmaleimide-resistant DNA polymerase activity was separated from the other three DNA polymerases by hydroxyapatite column chromatography. The enzyme, whose approximate sedimentation coefficient was 3.3S, effectively utilized poly(dA)-oligo(dT) as a template-primer and was completely inhibited by 10 μM 2', 3'-dideoxythymidine-5'-triphosphate but was not inhibited by aphidicolin. Therefore, this enzyme was classified as a eukaryotic DNA polymerase-β. Three species of the N-ethylmaleimide-sensitive DNA polymerase were separated with a phosphocellulose column. Two (5.5S and 6.5S) of the enzymes eluted from the column preferred activated DNA as a templateprimer. Both of the DNA polymerases were inhibited by aphidicolin but not by 2', 3'-dideoxythymidine-5'-triphosphate. Judging from the above results, these two DNA polymerases were classified as eukaryotic DNA polymerase-α. The third N-ethylmaleimide-sensitive enzyme (approximately 7S) eluted from the phospho-cellulose column utilized poly(rA)-oligo(dT) as well as poly(dA)-oligo(dT) as tem-plate-primers. The enzyme activity was resistant to aphidicolin but sensitive to 2', 3'-dideoxythymidine-5'-triphosphate. This enzyme resembles eukaryotic DNA polymerase-γ. Thus, it is concluded that the four species of DNA polymerase found in starfish oocytes resemble eukaryotic DNA polymerase-α, -β, and -γ.
- 社団法人 日本生化学会の論文
著者
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Haraguchi Tokuko
Department Of Physiological Chemistry And Nutrition Faculty Of Medicine University Of Tokyo
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Nagano Hiroshi
Department Of Cardiovascular Surgery Saitama Medical Center Jichi Medical University
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HARAGUCHI Tokuko
Department of Physiological Chemistry and Nutrition, Faculty of Medicine, The University of Tokyo
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