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A low molecular weight DNA polymerase from wheat embryos

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Abstract

The study of plant DNA polymerases lags far behind that concerning their animal or yeast counterpart. In this work we describe the first extensive purification to apparent homogeneity, as well as a detailed biochemical and immunological characterization, of a low molecular weight DNA polymerase (DNA polymerase CI) purified from wheat embryos. The monomeric enzyme is a basic protein having a molecular weight of 52 kDa. Polyclonal antibodies raised in rabbits against DNA polymerase CI did not inhibit animal DNA polymerases α and β or wheat DNA polymerase A, whereas wheat DNA polymerases CII and B were much less affected than the CI enzyme. Several properties of enzyme CI were studied. Some known inhibitors of DNA polymerase activity including aphidicolin, phosphonoacetic acid and heparin, did not affect DNA polymerase CI while the activity of this enzyme was strongly inhibited by ddTTP and N-ethylmaleimide. The polyamine spermine decreased markedly the enzyme activity, while spermidine produced a strong stimulation at the same concentrations that spermine inhibited the enzyme. The best template for this enzyme is poly dA-oligo dT, although polymerase CI can recognize significantly some synthetic polyribonucleotide templates (poly rC-oligo dG, poly rA-oligo dT) but only at a given protein/template primer ratio. The enzyme is blocked at the amino terminus, thus preventing the automatic sequencing of the protein. The amino acid analysis showed a striking similarity with the animal low molecular weight DNA polymerase β. The latter observation, as well as the effect of inhibitors (except N-ethylmaleimide which does not inhibit the animal polymerase) indicate that the DNA polymerase described in this work is a plant DNA polymerase very similar to the low molecular weight animal DNA polymerase β, an enzyme believed to be involved in nuclear DNA repair.

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Author notes

  1. Marie-Thérèse Gatius

    Present address: INRA-Laboratoire L.M.T.C., BP 527, 44026, Nantes Cedex 03, France

Authors and Affiliations

  1. Institut de Biochimie Cellulaire et Neurochimie du CNRS, 1 rue Camille Saint Saëns, 33077, Bordeaux Cedex, France

    Michel Castroviejo, Marie-Thérèse Gatius & Simon Litvak

Authors
  1. Michel Castroviejo
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  2. Marie-Thérèse Gatius
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  3. Simon Litvak
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Castroviejo, M., Gatius, MT. & Litvak, S. A low molecular weight DNA polymerase from wheat embryos. Plant Mol Biol 15, 383–397 (1990). https://doi.org/10.1007/BF00019156

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  • DOI: https://doi.org/10.1007/BF00019156

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