High molecular weight deoxyribonucleic acid polymerase of LF hepatoma. Purification and properties.
A high molecular weight DNA polymerase has been purified from the cytosol of a fast growing hepatoma: LF hepatoma. This enzyme sediments at 11.3 S under polymerization reaction conditions (6 mM KCl) and at 8.3 S in higher salt concentrations (200 mM KCl). In either case, no activity is seen in the 3 to 4 S region where low molecular weight DNA polymerase is found. The purified enzyme has a neutral pH optimum and requires a divalent cation, all four deoxyribonucleoside triphosphates and an initiated DNA template for maximal activity. The synthetic template specificity of LF DNA polymerase has been studied. Although this enzyme cannot copy a polyribonucleotide template, the ribostrand of a synthetic hybrid can be used with low efficiency as an initiator for the synthesis of the complementary deoxyribonucleotide strand. The activity of the purified enzyme is strongly inhibited by thiol-blocking agents. The general properties of LF DNA polymerase are similar to those of high molecular weight mammalian DNA polymerases. In our experimental conditions, the error frequency of this tumoral DNA polymerase was no greater than that  made by the purified high molecular weight DNA polymerase of regenerating rat liver.
D000818:Animals; D006528:Carcinoma, Hepatocellular / Q000201:enzymology*; D002499:Centrifugation, Density Gradient; D002846:Chromatography, Affinity; D003600:Cytosol / Q000201:enzymology; D004254:DNA Nucleotidyltransferases / Q000302:isolation & purification* / Q000378:metabolism; D004789:Enzyme Activation / Q000187:drug effects; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D008113:Liver Neoplasms; D008274:Magnesium / Q000494:pharmacology; D008345:Manganese / Q000494:pharmacology; D008970:Molecular Weight; D009374:Neoplasms, Experimental / Q000201:enzymology; D011188:Potassium / Q000494:pharmacology; D051381:Rats; D013698:Templates, Genetic
D008345:Manganese; D004254:DNA Nucleotidyltransferases; D008274:Magnesium; D011188:Potassium