Abstract
The eukaryotic mariner transposons are currently thought to have no sequence specificity for integration other than to insert within a TA contained in a degenerated [TA]1–4 tract, either in vitro or in vivo. We have investigated the properties of a suspected hotspot for the integration of the mariner Mos1 element, namely the Tn9 cat gene that encodes a chloramphenicol acetyl transferase. Using in vitro and bacterial transposition assays, we confirmed that the cat gene is a preferential target for MOS1 integration, whatever its sequence environment, copy number or chromosomal locus. We also observed that its presence increases transposition rates both in vitro and in bacterial assays. The structural and sequence features that constitute the attractiveness of cat were also investigated. We first demonstrated that supercoiling is essential for the cat gene to be a hot spot. In contrast to the situation for Tc1-like elements, DNA curvature and bendability were not found to affect integration target preferences. We found that Mos1 integrations do not occur randomly along the cat gene. All TA dinucleotides that are preferred for integration were found within either TATA or TA×TA motifs. However, these motifs are not sufficient to constitute an attractive dinucleotide, since four TATA and TA×TA sites are cold spots.
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Acknowledgments
This work was supported by the University of François Rabelais of Tours, and funded by grants from the European Commission (Project SyntheGeneDelivery, No 018716), the CNRS, the French Ministère de l’Education Nationale, de la Recherche et de la Technologie (MENRT), the Association Française contre la Myopathie, the Groupement de Recherche CNRS 2157. G. Crénès holds a doctoral fellowship from the European Commission, and S. Dion from the French MENRT. We would like to thank Dr. M. Chandler for providing chloramphenicol-mediated resistance strains, Dr. G. Demarre for providing the pSW29 plasmid, and Dr. L. Segalat providing the sequences of insertion in C. elegans rDNA. The English text has been revised by Dr. M. Ghosh.
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Communicated by D. Andersson.
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Crénès, G., Ivo, D., Hérisson, J. et al. The bacterial Tn9 chloramphenicol resistance gene: an attractive DNA segment for Mos1 mariner insertions. Mol Genet Genomics 281, 315–328 (2009). https://doi.org/10.1007/s00438-008-0414-6
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DOI: https://doi.org/10.1007/s00438-008-0414-6