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. 1979 Oct 10;7(3):715–726. doi: 10.1093/nar/7.3.715

Unique requirements for template primers of DNA polymerase beta from rat ascites hepatoma AH130 cells.

K Ono, A Ohashi, K Tanabe, A Matsukage, M Nishizawa, T Takahashi
PMCID: PMC328050  PMID: 503843

Abstract

The optimal condition for the rat DNA polymerase beta activity with (rA)n . (dT)12-18 as a template-primer was determined. The activity was remarkably affected by the concentration of the primer, (dT)12-18' and the mixing ratio of (dT)12-18 to (rA)n. DNA polymerase beta requires higher primer concentration (Km = 11.1 microM with respect to 3'-OH of the primer) than DNA polymerase gamma (Km = 0.04 microM) or oncornaviral DNA polymerase (Km = 0.08 microM) and the enzyme represented the maximum activity in the base ratio of 2:1 with (dT)12-18 and (rA)n suggesting the difference in reaction mechanisms of these enzymes. Under the optimized conditions, the specific activity of the near homogeneous preparation of DNA polymerase beta was 1,000,000 units per mg protein.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Bollum F. J. Mammalian DNA polymerases. Prog Nucleic Acid Res Mol Biol. 1975;15(0):109–144. doi: 10.1016/s0079-6603(08)60118-x. [DOI] [PubMed] [Google Scholar]
  2. Chang L. M., Bollum F. J. Low molecular weight deoxyribonucleic acid polymerase from rabbit bone marrow. Biochemistry. 1972 Mar 28;11(7):1264–1272. doi: 10.1021/bi00757a023. [DOI] [PubMed] [Google Scholar]
  3. Chang L. M. Low molecular weight deoxyribonucleic acid polymerase from calf thymus chromatin. I. Preparation of homogeneous enzyme. J Biol Chem. 1973 Jun 10;248(11):3789–3795. [PubMed] [Google Scholar]
  4. Chang L. M. The distributive nature of enzymatic DNA synthesis. J Mol Biol. 1975 Apr 5;93(2):219–235. doi: 10.1016/0022-2836(75)90129-1. [DOI] [PubMed] [Google Scholar]
  5. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  6. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  7. Lindell T. J., Weinberg F., Morris P. W., Roeder R. G., Rutter W. J. Specific inhibition of nuclear RNA polymerase II by alpha-amanitin. Science. 1970 Oct 23;170(3956):447–449. doi: 10.1126/science.170.3956.447. [DOI] [PubMed] [Google Scholar]
  8. Matsukage A., Bohn E. W., Wilson S. H. Differential sensitivity of low molecular weight DNA polymerase to sulfhydryl-blocking reagents. Biochim Biophys Acta. 1975 Mar 21;383(3):338–343. doi: 10.1016/0005-2787(75)90062-3. [DOI] [PubMed] [Google Scholar]
  9. Matsukage A., Bohn E. W., Wilson S. H. On the DNA polymerase III of mouse myeloma: partial purification and characterization. Biochemistry. 1975 Mar 11;14(5):1006–1020. doi: 10.1021/bi00676a020. [DOI] [PubMed] [Google Scholar]
  10. Matsukage A., Nishizawa M., Takahashi T. Difference in the mechanisms of poly(dT) synthesis by DNA polymerases beta and gamma. J Biochem. 1979 Jun;85(6):1551–1554. doi: 10.1093/oxfordjournals.jbchem.a132487. [DOI] [PubMed] [Google Scholar]
  11. Nakajima K., Ono K., Ito Y. Interconversion of molecular size of the DNA polymerase from Rauscher leukemia virus. Intervirology. 1974;3(5-6):332–341. doi: 10.1159/000149770. [DOI] [PubMed] [Google Scholar]
  12. Tanabe K., Takahashi T. Conversion of DNA polymerase extracted from rat ascites hepatoma cells. Biochem Biophys Res Commun. 1973 Jul 2;53(1):295–301. doi: 10.1016/0006-291x(73)91433-2. [DOI] [PubMed] [Google Scholar]
  13. Tanabe K., Takahashi T. Studies on the molecular species of DNA polymerase extracted from rat ascites hepatoma cells. J Biochem. 1976 Jan;79(1):85–90. doi: 10.1093/oxfordjournals.jbchem.a131061. [DOI] [PubMed] [Google Scholar]
  14. Wang T. S., Sedwick W. D., Korn D. Nuclear deoxyribonucleic acid polymerase. Further observations on the structure and properties of the enzyme from human KB cells. J Biol Chem. 1975 Sep 10;250(17):7040–7044. [PubMed] [Google Scholar]

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