Dissociation and reconstitution of a DNA polymerase alpha-primase complex

J Biochem. 1985 Aug;98(2):581-4. doi: 10.1093/oxfordjournals.jbchem.a135314.


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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cell Line
  • DNA Polymerase II / isolation & purification
  • DNA Polymerase II / metabolism*
  • DNA Primase
  • Ethylene Glycol
  • Ethylene Glycols / pharmacology
  • Kinetics
  • Mice
  • Molecular Weight
  • Multienzyme Complexes / isolation & purification
  • Multienzyme Complexes / metabolism*
  • RNA Nucleotidyltransferases / isolation & purification
  • RNA Nucleotidyltransferases / metabolism*
  • Teratoma


  • Ethylene Glycols
  • Multienzyme Complexes
  • DNA Primase
  • RNA Nucleotidyltransferases
  • DNA Polymerase II
  • Ethylene Glycol