Dissociation and reconstitution of a DNA polymerase alpha-primase complex.
The conditions for dissociation of the DNA polymerase alpha-primase complex (DNA polymerase alpha 1) have been examined. It was revealed that 50% ethylene glycol effectively dissociated the complex. The dissociated DNA polymerase and primase were purified to eliminate cross-contaminating activities by column chromatography using buffers containing 50% ethylene glycol. The sedimentation coefficients of the purified DNA polymerase and primase were 7.1S and 5.7S, respectively. These two enzymes were mixed in the presence of 20% ethylene glycol and the mixture was sedimented through a glycerol gradient containing no ethylene glycol. The DNA polymerase and primase activities co-sedimented at 9.1S which corresponds to the S value of intact alpha 1, indicating the reconstitution of the DNA polymerase alpha-primase complex.
D000818:Animals; D002460:Cell Line; D004257:DNA Polymerase II / Q000302:isolation & purification / Q000378:metabolism*; D019915:DNA Primase; D019855:Ethylene Glycol; D005026:Ethylene Glycols / Q000494:pharmacology; D007700:Kinetics; D051379:Mice; D008970:Molecular Weight; D009097:Multienzyme Complexes / Q000302:isolation & purification / Q000378:metabolism*; D012316:RNA Nucleotidyltransferases / Q000302:isolation & purification / Q000378:metabolism*; D013724:Teratoma
D005026:Ethylene Glycols; D009097:Multienzyme Complexes; D019915:DNA Primase; D012316:RNA Nucleotidyltransferases; D004257:DNA Polymerase II; D019855:Ethylene Glycol