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Novikoff hepatoma deoxyribonucleic acid polymerase. Sensitivity of the beta-polymerase to sulfhydryl blocking agents.

Publication ,  Journal Article
Mosbaugh, DW; Kunkel, TA; Stalker, DM; Tcheng, JE; Meyer, RR
Published in: Nucleic Acids Res
September 1976

Unlike other beta-class eukaryotic DNA polymerases, the enzyme purified from the Novikoff hepatoma is inhibited by both sulfhydryl blocking agents N-ethylmaleimide (NEM) and p-hydroxymercuribenzoate (pHMB). The degree of sensitivity varies depending on the enzyme purity, pH of the reaction, and the presence of sulfhydryl reducing agents. Novikoff beta-polymerase activity is unaffected by the presence of 2-mercaptoethanol (2-Me) or dithiothreitol (DTT); however, the combination of 2-mercaptoethanol and NEM or pHMB acts to reverse the inhibition of the sulfhydryl blocking agent. The reversal of inhibition involves more than just a titration of NEM with 2-mercaptoethanol since a) the combination of these two reagents actually stimulates the DNA polymerase, and b) dithiothreitol did not reverse the inhibition. Binding of the polymerase to DNA did not affect the enzyme sensitivity to NEM.

Duke Scholars

Published In

Nucleic Acids Res

DOI

ISSN

0305-1048

Publication Date

September 1976

Volume

3

Issue

9

Start / End Page

2341 / 2352

Location

England

Related Subject Headings

  • Sulfhydryl Reagents
  • Sea Urchins
  • Neoplasms, Experimental
  • Mercaptoethanol
  • Liver Neoplasms
  • Liver
  • Hydroxymercuribenzoates
  • Hydrogen-Ion Concentration
  • Guinea Pigs
  • Ethylmaleimide
 

Citation

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Mosbaugh, D. W., Kunkel, T. A., Stalker, D. M., Tcheng, J. E., & Meyer, R. R. (1976). Novikoff hepatoma deoxyribonucleic acid polymerase. Sensitivity of the beta-polymerase to sulfhydryl blocking agents. Nucleic Acids Res, 3(9), 2341–2352. https://doi.org/10.1093/nar/3.9.2341
Mosbaugh, D. W., T. A. Kunkel, D. M. Stalker, J. E. Tcheng, and R. R. Meyer. “Novikoff hepatoma deoxyribonucleic acid polymerase. Sensitivity of the beta-polymerase to sulfhydryl blocking agents.Nucleic Acids Res 3, no. 9 (September 1976): 2341–52. https://doi.org/10.1093/nar/3.9.2341.
Mosbaugh DW, Kunkel TA, Stalker DM, Tcheng JE, Meyer RR. Novikoff hepatoma deoxyribonucleic acid polymerase. Sensitivity of the beta-polymerase to sulfhydryl blocking agents. Nucleic Acids Res. 1976 Sep;3(9):2341–52.
Mosbaugh, D. W., et al. “Novikoff hepatoma deoxyribonucleic acid polymerase. Sensitivity of the beta-polymerase to sulfhydryl blocking agents.Nucleic Acids Res, vol. 3, no. 9, Sept. 1976, pp. 2341–52. Pubmed, doi:10.1093/nar/3.9.2341.
Mosbaugh DW, Kunkel TA, Stalker DM, Tcheng JE, Meyer RR. Novikoff hepatoma deoxyribonucleic acid polymerase. Sensitivity of the beta-polymerase to sulfhydryl blocking agents. Nucleic Acids Res. 1976 Sep;3(9):2341–2352.
Journal cover image

Published In

Nucleic Acids Res

DOI

ISSN

0305-1048

Publication Date

September 1976

Volume

3

Issue

9

Start / End Page

2341 / 2352

Location

England

Related Subject Headings

  • Sulfhydryl Reagents
  • Sea Urchins
  • Neoplasms, Experimental
  • Mercaptoethanol
  • Liver Neoplasms
  • Liver
  • Hydroxymercuribenzoates
  • Hydrogen-Ion Concentration
  • Guinea Pigs
  • Ethylmaleimide