Size difference in catalytic polypeptides of two active forms of mouse DNA polymerase alpha and separation of the primase subunit from one form, DNA replicase.
There are two active forms of DNA polymerase alpha in mouse cells. One form (DNA replicase) is a DNA polymerase associated with primase activity and the other form (7.3 S polymerase) has no primase activity (Yaugar, T., Kozu, T. and Seno, T. (1982) J. Biol. Chem. 257, 11121-11127). The primase activity was dissociated from partially purified DNA replicase by hydroxyapatite column chromatography in buffer containing dimethyl sulfoxide and ethylene glycol. Nearly homogeneous primase, consisting of a 58 kDa polypeptide was obtained by glycerol gradient sedimentation and DEAE-cellulose column chromatography. Experiments on the effect of proteinase treatment and measurement of the molecular weight of the catalytic polypeptide of DNA replicase after its dissociation from the primase polypeptide indicated that the primase is not part of the DNA polymerase molecule, but an independent protein associated with DNA polymerase alpha, and that the latter is a 115 kDa catalytic polypeptide. The other form of DNA polymerase alpha, 7.3 S polymerase, consists of a 72 kDa catalytic polypeptide. Thus, the two forms of mouse DNA polymerase alpha have partially, if not completely, different catalytic polypeptide structures, suggesting that the 7.3 S polymerase is not simply formed from DNA replicase by dissociation of the primase subunit.
D000818:Animals; D002286:Carcinoma, Ehrlich Tumor / Q000032:analysis; D002848:Chromatography, DEAE-Cellulose; D004257:DNA Polymerase II / Q000032:analysis*; D019915:DNA Primase; D004259:DNA-Directed DNA Polymerase / Q000032:analysis*; D004591:Electrophoresis, Polyacrylamide Gel; D010450:Endopeptidases / Q000494:pharmacology; D051379:Mice; D008970:Molecular Weight; D009363:Neoplasm Proteins / Q000032:analysis; D010455:Peptides / Q000032:analysis; D012316:RNA Nucleotidyltransferases / Q000037:antagonists & inhibitors / Q000302:isolation & purification*
D009363:Neoplasm Proteins; D010455:Peptides; D019915:DNA Primase; C023140:DNA replicase; D012316:RNA Nucleotidyltransferases; D004257:DNA Polymerase II; D004259:DNA-Directed DNA Polymerase; D010450:Endopeptidases