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Recognition and processing of the origin of transfer DNA by conjugative relaxase TrwC

Nature Structural & Molecular Biology volume 10, pages 1002–1010 (2003)Cite this article

A Corrigendum to this article was published on 01 January 2004

Abstract

Relaxases are DNA strand transferases that catalyze the initial and final stages of DNA processing during conjugative cell-to-cell DNA transfer. Upon binding to the origin of transfer (oriT) DNA, relaxase TrwC melts the double helix. The three-dimensional structure of the relaxase domain of TrwC in complex with its cognate DNA at oriT shows a fold built on a two-layer α/β sandwich, with a deep narrow cleft that houses the active site. The DNA includes one arm of an extruded cruciform, an essential feature for specific recognition. This arm is firmly embraced by the protein through a β-ribbon positioned in the DNA major groove and a loop occupying the minor groove. It is followed by a single-stranded DNA segment that enters the active site, after a sharp U-turn forming a hydrophobic cage that traps the N-terminal methionine. Structural analysis combined with site-directed mutagenesis defines the architecture of the active site.

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Figure 1: Sequence alignment and scheme of DNA processing by TrwC.
Figure 2: TrwC binding to oriT DNA.
Figure 3: Structure of TrwC-N293 in complex with DNA.
Figure 4: Protein-DNA interactions.
Figure 5: Details of TrwC-DNA interactions.
Figure 6: Electron density map and active site.

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Acknowledgements

This study was supported by the Spain Ministerio de Educación y Ciencia, the Generalitat de Catalunya and the European Union (EU). Synchrotron data collection was supported by the European Synchrotron Radiation Facility (ESRF) and the EU. We thank A.G. Blanco for generating Figure 1b and helping with data collection.

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Authors and Affiliations

  1. Institut de Biologia Molecular de Barcelona, CSIC, Jordi Girona, 18-26, Barcelona, 08034, Spain

    Alicia Guasch, Rosa Pérez-Luque, F Xavier Gomis-Rüth & Miquel Coll

  2. Departamento de Biología Molecular (Laboratorio asociado al CIB, CSIC), Universidad de Cantabria, Herrera Oria, s/n, Santander, 39011, Spain

    María Lucas, Gabriel Moncalián, Matilde Cabezas & Fernando de la Cruz

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  1. Alicia Guasch
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Correspondence to Fernando de la Cruz or Miquel Coll.

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Guasch, A., Lucas, M., Moncalián, G. et al. Recognition and processing of the origin of transfer DNA by conjugative relaxase TrwC. Nat Struct Mol Biol 10, 1002–1010 (2003). https://doi.org/10.1038/nsb1017

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