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Molecular genetic and biochemical analysis of Brassica napus proliferating cell nuclear antigen function

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Abstract

A cDNA encoding the proliferating cell nuclear antigen (PCNA) from Brassica napus (oilseed rape) was shown to complement the lethal deletion mutation in the PCNA gene (ΔPOL30) of Saccharomyces cerevisiae. We provide unequivocal evidence that the B. napus PCNA can perform all the essential functions of the yeast PCNA in DNA replication, although some species-specific differences may exist. In addition, the B. napus PCNA expressed as a fusion polypeptide with glutathione S-transferase (GST) was shown to stimulate the activity and processivity of two δ-like DNA polymerases from wheat in vitro. These experiments provide direct biochemical evidence that the B. napus PCNA may function as an auxiliary factor in plant cell DNA replication.

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Authors and Affiliations

  1. Department of Biological Sciences, University of Calgary, 2500 University Drive, NW, Calgary, Alberta, Canada, T2N 1N4

    Maurice M. Moloney

  2. Department of Medical Biochemistry, University of Calgary Health Sciences Center, Calgary, Alberta, Canada, T2N 4N1

    Nancy-Ann Markley & Dallan Young

  3. Institut de Biochimie et Génétique Cellulaires du Centre National de la Recherche Scientifique, Bordeaux, France

    Patricia Laquel & Michel Castroviejo

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  1. Nancy-Ann Markley
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  2. Dallan Young
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  3. Patricia Laquel
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  4. Michel Castroviejo
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  5. Maurice M. Moloney
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Markley, NA., Young, D., Laquel, P. et al. Molecular genetic and biochemical analysis of Brassica napus proliferating cell nuclear antigen function. Plant Mol Biol 34, 693–700 (1997). https://doi.org/10.1023/A:1005817220344

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