PMID: 974129Oct 18, 1976

Characterization of DNA polymerase associated with nuclear membrane fractions from uninfected and adenovirus 2-infected KB cells

Biochimica Et Biophysica Acta
K ItoM GREEN

Abstract

We have isolated a nuclear membrane fraction from KB cells infected with human adenovirus 2 that synthesizes exclusively small viral DNA chains (approx. 9 S) in vitro (Yamashita, T., Arens, M. and Green, M. (1975) J. Biol. Chem. 250, 3273-3279). The DNA polymerase activity present in the adenovirus 2 DNA-nuclear membrane complex was purified through chromatography on phosphocellulose and DEAE-cellulose, glycerol gradient centrifuation and DNA-cellulose chromatography. A single peak of enzymatic activity sedimented in glycerol gradients at about 6.7 S which corresponds to a molecular weight of 125000. The enzyme preparation in the step of glycerol gradient centrifugation utilized activated calf thymus, KB cell and adenovirus 2 DNA as template-primer in the presence of Mg2+; Km values for these DNAs were 5.5, 4.0, and 0.8 mug/ml, respectively. The partially purified enzyme preparation was characterized by several criteria which were compared to the properties of the three major mammalian DNA polymerases, alpha, beta, and psi. On the basis of template-primer preference, effect of salt, inhibition by N-ethylmaleimide and Km for dTTP, the DNA polymerase activity from the membrane complex can be distinguished from the alpha and beta ...Continue Reading

References

Feb 1, 1975·Proceedings of the National Academy of Sciences of the United States of America·S Spadari, A Weissbach
Nov 1, 1972·Archives of Biochemistry and Biophysics·A BoldenA Weissbach
Feb 1, 1973·Proceedings of the National Academy of Sciences of the United States of America·R McCaffreyD Baltimore
Feb 1, 1972·Proceedings of the National Academy of Sciences of the United States of America·B FridlenderA Weissbach
Jan 1, 1962·Cold Spring Harbor Symposia on Quantitative Biology·M GREEN
May 15, 2009·Chemistry : a European Journal·Eduardo J FernándezEva Sánchez-Forcada

Citations

Jul 31, 1990·Biochemical and Biophysical Research Communications·M Goulian, S L Grimm
Mar 8, 2012·Journal of Cellular Biochemistry·Rebeca C Castillo-Mora, Armando Aranda-Anzaldo
Apr 16, 2019·Journal of Neuroscience Research·Myrna A R Dent, Armando Aranda-Anzaldo

Related Concepts

Adenoviruses, Human
Protoplasm
DNA Polymerase N3
DNA, Single-Stranded
DNA, Viral
Endonuclease
Ethylmaleimide
Magnesium
Manganese
Nuclear Envelope

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