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Repeat units from a maize rDNA external spacer region exhibit DNA curvature and interact with high-mobility-group proteins

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Abstract

The 3-kb external spacer from a maize (Zea mays L. cv. A619) nuclear rRNA gene unit which contains nine highly homologous 200-bp repeat elements was found to include a region with DNA-curvature properties. The centre of curvature was localized within repeats 5 and 6 using a circular permutation assay. A 60-bp-long subfragment of this region was found to interact with nuclear proteins, including high-mobility-group (HMG) proteins, and with the maize HMGa protein synthesized in Escherichia coli from a recombinant plasmid. The potential influence of the binding of the HMG proteins on the conformation of this subfragment was studied with a permutation assay based on a bending vector.

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Abbreviations

bp:

base pairs

ESR:

external spacer region

HMG:

high mobility group

Pol I:

RNA polymerase I

EMSA:

electrophoretic mobility-shift assay

TCA:

trichloroacetic acid

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We thank Dr. S. Adhya (Laboratory of Molecular Biology, National Cancer Institute, Bethesda, USA) for plasmid pBend2 and Dr. U.-G. Maier (Institut für Biologie II, Albert-Ludwigs-Universität Freiburg, FRG) for a subclone of pMS1. Dr. G. Igloi (Institut für Biologie III, Albert-Ludwigs-Universität Freiburg, FRG) and Dr. T. Quayle (Department of Biological Sciences, University of Wisconsin-Parkside, Wis., USA) are thanked for comments on the manuscript, and Hannelore Jonas for excellent technical assistance. This work was supported by grants from the Deutsche Forschungsgemeinschaft and by the Fond der Chemischen Industrie.

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Griess, E.A., Grasser, K.D. & Feix, G. Repeat units from a maize rDNA external spacer region exhibit DNA curvature and interact with high-mobility-group proteins. Planta 191, 524–531 (1993). https://doi.org/10.1007/BF00195754

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  • DOI: https://doi.org/10.1007/BF00195754

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