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Characterization of topoisomerase I and II activities in nuclear extracts during callogenesis in immature embryos of Zea mays

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Abstract

We have characterized the topoisomerase I and II activities in nuclear extracts from immature embryos of Zea mays and the effect of the treatment with 2,4-dichlorophenoxyacetic acid (2,4-D) and abscisic acid (ABA). These extracts were shown to be essentially devoid of protease and nuclease activities and they were tested for their ability to relax supercoiled DNA, unknotting P4 DNA and catenate circular duplex DNA under catalytic conditions. Unknotting and catenation reactions are strictly magnesium- and ATP-dependent, but not the relaxation of circular supercoiled DNA allowing the detection of both topoisomerase I and II activities. Two cytotoxic drugs, camptothecin, a plant alkaloid that inhibits cukaryotic topoisomerase I, and epipodophyllotoxin VM-26 (teniposide) that inhibits topoisomerase II, have been assayed in our extracts showing similar inhibitory effects on topoisomerase enzymes. Alkaline phosphatase treatment of nuclear extracts abolishes both topoisomerase activities. Nuclear extracts from embryos treated with 2,4-D showed 200% increase on topoisomerase II activity as compared with untreated ones, but only residual activity was detected in ABA-treated embryos. Nuclear extracts from hormone-treated and untreated embryos showed similar topoisomerase I activity with deviations of less than 25%. These differences are discussed in terms of possible post-translational modifications of the enzymes associated with the increase in proliferation activity of calli.

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Authors and Affiliations

  1. Departmento de Genética Molecular, Centro de Investigación y Desarrollo, CSIC, Jordi Girona 18, 08034, Barcelona, Spain

    Miguel Carballo, Ramon Giné & Pere Puigdomènech

  2. Departamento de Biología Molecular y Agrobiología, Centro de Investigación y Desarrollo, CSIC, Jordi Girona 18, 08034, Barcelona, Spain

    Mireia Santos

Authors
  1. Miguel Carballo
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  2. Ramon Giné
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  3. Mireia Santos
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  4. Pere Puigdomènech
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Carballo, M., Giné, R., Santos, M. et al. Characterization of topoisomerase I and II activities in nuclear extracts during callogenesis in immature embryos of Zea mays . Plant Mol Biol 16, 59–70 (1991). https://doi.org/10.1007/BF00017917

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  • DOI: https://doi.org/10.1007/BF00017917

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