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Nucleic Acids Research Advance Access published online on November 13, 2007
Nucleic Acids Research, doi:10.1093/nar/gkm759
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© 2007 The Author(s)
This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/2.0/uk/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

Molecular Biology

Dynamic structures of Bacillus subtilis RecN–DNA complexes

Humberto Sanchez1,2, Paula P. Cardenas1, Shige H. Yoshimura2, Kunio Takeyasu2 and Juan C. Alonso1,*

1Departamento de Biotecnología Microbiana, Centro Nacional de Biotecnología, CSIC, Campus Universidad Autónoma de Madrid, Darwin 3, Cantoblanco, 28049 Madrid, Spain and 2Laboratory of Plasma Membrane and Nuclear Signalling, Graduate School of Biostudies, Kyoto University, Sakyo-ku, Kitashirakawa-Oiwake-cho, Kyoto 606-8502, Japan

*To whom correspondence should be addressed. Tel: +34 91585 4546; Fax: +34 91585 4506; Email: jcalonso{at}cnb.uam.es

Received July 20, 2007. Revised September 10, 2007. Accepted September 11, 2007.

Genetic and cytological evidences suggest that Bacillus subtilis RecN acts prior to and after end-processing of DNA double-strand ends via homologous recombination, appears to participate in the assembly of a DNA repair centre and interacts with incoming single-stranded (ss) DNA during natural transformation. We have determined the architecture of RecN–ssDNA complexes by atomic force microscopy (AFM). ATP induces changes in the architecture of the RecN–ssDNA complexes and stimulates inter-complex assembly, thereby increasing the local concentration of DNA ends. The large CII and CIII complexes formed are insensitive to SsbA (counterpart of Escherichia coli SSB or eukaryotic RPA protein) addition, but RecA induces dislodging of RecN from the overhangs of duplex DNA molecules. Reciprocally, in the presence of RecN, RecA does not form large RecA–DNA networks. Based on these results, we hypothesize that in the presence of ATP, RecN tethers the 3'-ssDNA ends, and facilitates the access of RecA to the high local concentration of DNA ends. Then, the resulting RecA nucleoprotein filaments, on different ssDNA segments, might promote the simultaneous genome-wide homology search.


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