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Título: | Bacillus subtilis MutS Modulates RecA-Mediated DNA Strand Exchange Between Divergent DNA Sequences. |
Autor: | Carrasco, Begoña; Serrano, E.; Martín-González, Alejandro; Moreno-Herrero, Fernando; Alonso, Juan Carlos CSIC ORCID | Palabras clave: | MutS RecA nucleoprotein filaments SsbA Genetic variation Horizontal gene transfer Mismatch repair |
Fecha de publicación: | feb-2019 | Editor: | Frontiers Media | Citación: | Frontiers in microbiology 10: 237 (2019) | Resumen: | The efficiency of horizontal gene transfer, which contributes to acquisition and spread of antibiotic resistance and pathogenicity traits, depends on nucleotide sequence and different mismatch-repair (MMR) proteins participate in this process. To study how MutL and MutS MMR proteins regulate recombination across species boundaries, we have studied natural chromosomal transformation with DNA up to ∼23% sequence divergence. We show that Bacillus subtilis natural chromosomal transformation decreased logarithmically with increased sequence divergence up to 15% in wild type (wt) cells or in cells lacking MutS2 or mismatch repair proteins (MutL, MutS or both). Beyond 15% sequence divergence, the chromosomal transformation efficiency is ∼100-fold higher in ΔmutS and ΔmutSL than in ΔmutS2 or wt cells. In the first phase of the biphasic curve (up to 15% sequence divergence), RecA-catalyzed DNA strand exchange contributes to the delineation of species, and in the second phase, homology-facilitated illegitimate recombination might aid in the restoration of inactivated genes. To understand how MutS modulates the integration process, we monitored DNA strand exchange reactions using a circular single-stranded DNA and a linear double-stranded DNA substrate with an internal 77-bp region with ∼16% or ∼54% sequence divergence in an otherwise homologous substrate. The former substrate delayed, whereas the latter halted RecA-mediated strand exchange. Interestingly, MutS addition overcame the heterologous barrier. We propose that MutS assists DNA strand exchange by facilitating RecA disassembly, and indirectly re-engagement with the homologous 5'-end of the linear duplex. Our data supports the idea that MutS modulates bidirectional RecA-mediated integration of divergent sequences and this is important for speciation. | Versión del editor: | https://doi.org/10.3389/fmicb.2019.00237 | URI: | http://hdl.handle.net/10261/185817 | DOI: | 10.3389/fmicb.2019.00237 | E-ISSN: | 1664-302X |
Aparece en las colecciones: | (CNB) Artículos |
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Frontiers in Microbiol. 2019.pdf | Artículo principal | 5,21 MB | Adobe PDF | Visualizar/Abrir |
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