Purification and properties of a low molecular weight DNA polymerase from Neurospora crassa.
A third DNA polymerase 'C' with low molecular weight was isolated and purified 3700-fold from ground hyphae of Neurospora crassa WT 74 A, which shows similarities to beta- and gamma-polymerases from higher eukaryotes: preference for poly(rA)(dT) as a template/primer, inhibition by p-chloromercuribenzoate, resistance against N-ethylmaleimide up to 10 mmol/l, and molecular weight of about 40000. This polymerase elutes as a distinct peak from DEAE-cellulose at 0.60 mol/l KCl and has an optimum for K+ at 2-20 mmol/l, for Mn2+ at 0.8 mmol/l, for Mg2+ at 4.0 mmol/l, the pH optimum is 8.0. Its Km is 1.5 mumol/l using dTTP as substrate. The enzyme activity described here is free of endonuclease but contains detectable amounts of exonuclease.
D002846:Chromatography, Affinity; D007700:Kinetics; D008970:Molecular Weight; D009491:Neurospora / Q000201:enzymology*; D009492:Neurospora crassa / Q000201:enzymology*; D009713:Nucleotidyltransferases / Q000302:isolation & purification* / Q000378:metabolism; D012194:RNA-Directed DNA Polymerase / Q000378:metabolism; D013439:Sulfhydryl Reagents / Q000494:pharmacology; D013698:Templates, Genetic
D013439:Sulfhydryl Reagents; C040220:DNA polymerase C; D009713:Nucleotidyltransferases; D012194:RNA-Directed DNA Polymerase