title: Purification of Epstein-Barr virus DNA polymerase from P3HR-1 cells.
abstract: The Epstein-Barr virus DNA polymerase was purified from extracts of P3HR-1 cells treated with n-butyrate for induction of the viral cycle. Sequential chromatography on DNA cellulose, phosphocellulose, and blue Sepharose yielded an enzyme preparation purified more than 1,300-fold. The purified enzyme was distinct from cellular enzymes but resembled the viral DNA polymerase in cells infected with herpes simplex virus type 1 or 2. The active enzyme had an apparent molecular weight of 185,000 as estimated by gel filtration on Sephacryl S-300. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis revealed a major polypeptide corresponding to a molecular weight of ca. 110,000. This polypeptide correlated with the catalytic function of the purified enzyme, whereas the other, less abundant polypeptides did not. By immunoblotting, the 110,000-molecular-weight polypeptide could be identified as a viral polypeptide. It could not be determined whether the native enzyme was composed of more than one polypeptide.
authors.forename: B
authors.initials: B
authors.lastname: Kallin
authors.forename: L
authors.initials: L
authors.lastname: Sternås
authors.forename: A K
authors.initials: AK
authors.lastname: Saemundssen
authors.forename: J
authors.initials: J
authors.lastname: Luka
authors.forename: H
authors.initials: H
authors.lastname: Jörnvall
authors.forename: B
authors.initials: B
authors.lastname: Eriksson
authors.forename: P Z
authors.initials: PZ
authors.lastname: Tao
authors.forename: M T
authors.initials: MT
authors.lastname: Nilsson
authors.forename: G
authors.initials: G
authors.lastname: Klein
mesh_terms: D000818:Animals; D002087:Butyrates / Q000494:pharmacology; D020148:Butyric Acid; D004259:DNA-Directed DNA Polymerase / Q000032:analysis / Q000096:biosynthesis / Q000302:isolation & purification*; D004854:Herpesvirus 4, Human / Q000201:enzymology*; D008970:Molecular Weight; D018139:Simplexvirus / Q000201:enzymology
chemical_list: D002087:Butyrates; D020148:Butyric Acid; D004259:DNA-Directed DNA Polymerase