Purification and Properties of DNA Polymerase Gamma From Rabbit Intestinal Epithelial Cells

Acta Biochim Pol. 1979;26(4):335-47.

Abstract

1. DNA polymerase gamma from the cytoplasmic fraction of rabbit intestinal epithelial cells has been purified 120 000-fold and was free of phosphatase and nuclease activities towards deoxyribonucleoside-5'-triphosphates and polynucleotides. 2. The enzyme exhibited maximal activity for activated DNA and poly(A) . oligo(dT)12--18 at pH 8.5 IN 0.25 AND 0.15 M-KCl, respectively. Km values for dTTP with these two templates were 0.5 and 3.8 microM, respectively. 3. In contrast to DNA polymerases alpha and beta, the enzyme replicated poly(A) . oligo(dT)12--18 10 times faster and poly(dA) . oligo(dT)12--18 5 times slower than activated DNA. 4. DNA polymerase gamma did not replicate poly(C) . oligo(dG)12--18 or poly(Cm) . oligo(dT)12--18. The reaction with poly(I) and poly(U) did not exceed 1% of that observed with poly(A). 5. The enzyme was inhibited in 60% by antiserum against DNA polymerase gamma from human lymphoblasts. 6. The nuclear fraction of rabbit intestinal epithelial cells contained DNA polymerase gamma with the same characteristics.

MeSH terms

  • Animals
  • Cell Nucleus / enzymology
  • Cytoplasm / enzymology*
  • DNA Polymerase III / antagonists & inhibitors
  • DNA Polymerase III / isolation & purification*
  • DNA-Directed DNA Polymerase / isolation & purification*
  • Goats
  • Humans
  • Immune Sera
  • Immunoassay / methods
  • Immunoglobulin G
  • Intestine, Small / enzymology*
  • Intestine, Small / ultrastructure
  • Lymphocytes / immunology
  • Polyribonucleotides / biosynthesis
  • Rabbits

Substances

  • Immune Sera
  • Immunoglobulin G
  • Polyribonucleotides
  • DNA Polymerase III
  • DNA-Directed DNA Polymerase