Purification of Epstein-Barr virus DNA polymerase from P3HR-1 cells.
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.
Kallin
B
B
Sternås
L
L
Saemundssen
A K
AK
Luka
J
J
Jörnvall
H
H
Eriksson
B
B
Tao
P Z
PZ
Nilsson
M T
MT
Klein
G
G
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
D002087:Butyrates; D020148:Butyric Acid; D004259:DNA-Directed DNA Polymerase