Purification and properties of DNA polymerase gamma from rabbit intestinal epithelial cells.
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.
D000818:Animals; D002467:Cell Nucleus / Q000201:enzymology; D003593:Cytoplasm / Q000201:enzymology*; D004258:DNA Polymerase III / Q000037:antagonists & inhibitors / Q000302:isolation & purification*; D004259:DNA-Directed DNA Polymerase / Q000302:isolation & purification*; D006041:Goats; D006801:Humans; D007106:Immune Sera; D007118:Immunoassay / Q000379:methods; D007074:Immunoglobulin G; D007421:Intestine, Small / Q000201:enzymology* / Q000648:ultrastructure; D008214:Lymphocytes / Q000276:immunology; D011131:Polyribonucleotides / Q000096:biosynthesis; D011817:Rabbits
D007106:Immune Sera; D007074:Immunoglobulin G; D011131:Polyribonucleotides; D004258:DNA Polymerase III; D004259:DNA-Directed DNA Polymerase