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A NMR guided approach for CsrA–RNA crystallization

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Abstract

Structure determination of protein–nucleic acid complexes remains a challenging task. Here we present a simple method for generating crystals of a CsrA–nucleic acid complex, guided entirely by results from nuclear magnetic resonances spectroscopy (NMR) spectroscopy. Using a construct that lacks thirteen non-essential C-terminal residues, efficient binding to DNA could be demonstrated. One CsrA dimer interacts with two DNA oligonucleotides, similar to previous findings with RNA. Furthermore, the NMR study of the CsrA–DNA complex was the basis for successfully homing in on conditions that were suitable for obtaining crystals of the CsrA–DNA complex. Our results may be useful for those cases where RNA in protein–nucleic acid complexes may be replaced by DNA.

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Acknowledgments

The authors thank Mike Delk for NMR technical support.

Conflict of interest

The authors declare that they have no competing financial interests.

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Authors and Affiliations

  1. Department of Structural Biology, University of Pittsburgh School of Medicine, 3501 Fifth Avenue, BST3/Rm1050, Pittsburgh, PA, 15260, USA

    Leonardus M. I. Koharudin & Angela M. Gronenborn

  2. NMR Spectroscopy, Bijvoet Centrum for Biomolecular Research, Utrecht University, 3584 CH, Utrecht, The Netherlands

    Rolf Boelens & Rob Kaptein

  3. Novosibirsk State University, Pirogova 2, 630090, Novosibirsk, Russia

    Rob Kaptein

Authors
  1. Leonardus M. I. Koharudin
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  2. Rolf Boelens
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  3. Rob Kaptein
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  4. Angela M. Gronenborn
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Correspondence to Angela M. Gronenborn.

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Koharudin, L.M.I., Boelens, R., Kaptein, R. et al. A NMR guided approach for CsrA–RNA crystallization. J Biomol NMR 56, 31–39 (2013). https://doi.org/10.1007/s10858-013-9712-3

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  • DOI: https://doi.org/10.1007/s10858-013-9712-3

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