title: Purification and characterization of a 100 kDa DNA polymerase from cauliflower inflorescence.
abstract: A DNA polymerase from cauliflower (Brassica oleracea var. botrytis) inflorescence has been purified to near homogeneity through five successive column chromatographies, and temporally designated cauliflower polymerase 1. Cauliflower polymerase 1 is a monopolypeptide with a molecular mass of 100 kDa. The enzyme efficiently uses synthetic DNA homopolymers and moderately activated DNA and a synthetic RNA homopolymer as template-primers. The enzyme is strongly sensitive to dideoxythymidine triphosphate and N-ethylmaleimide, but it is insensitive to aphidicolin. It was stimulated with 250 mM KCl. Its mode of DNA synthesis is high-processive with or without proliferating-cell nuclear antigen. A 3'-->5' exonuclease activity is associated with cauliflower polymerase 1. The enzyme is clearly different from cauliflower mitochondrial polymerase and does not resemble the four different types of wheat DNA polymerase, designated wheat DNA polymerases A, B, CI and CII. In the present paper the role of the enzyme in plant DNA synthesis is discussed.
authors.affiliation: Department of Applied Biological Science, Faculty of Science and Technology, Science University of Tokyo, 2641 Yamazaki, Noda-shi, Chiba-ken 278, Japan.
authors.forename: H
authors.initials: H
authors.lastname: Seto
authors.forename: M
authors.initials: M
authors.lastname: Hatanaka
authors.forename: S
authors.initials: S
authors.lastname: Kimura
authors.forename: M
authors.initials: M
authors.lastname: Oshige
authors.forename: Y
authors.initials: Y
authors.lastname: Tsuya
authors.forename: Y
authors.initials: Y
authors.lastname: Mizushina
authors.forename: T
authors.initials: T
authors.lastname: Sawado
authors.forename: N
authors.initials: N
authors.lastname: Aoyagi
authors.forename: T
authors.initials: T
authors.lastname: Matsumoto
authors.forename: J
authors.initials: J
authors.lastname: Hashimoto
authors.forename: K
authors.initials: K
authors.lastname: Sakaguchi
mesh_terms: D001937:Brassica / Q000201:enzymology*; D002412:Cations / Q000494:pharmacology; D004259:DNA-Directed DNA Polymerase / Q000737:chemistry*; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005092:Exonucleases / Q000378:metabolism; D008970:Molecular Weight; D010940:Plant Proteins / Q000737:chemistry; D013379:Substrate Specificity
chemical_list: D002412:Cations; D004791:Enzyme Inhibitors; D010940:Plant Proteins; D004259:DNA-Directed DNA Polymerase; D005092:Exonucleases