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Title

Discovery of a new family of relaxases in Firmicutes bacteria

AuthorsRamachandran, Gayetri CSIC; Miguel-Arribas, Andrés; Abia, David CSIC ORCID ; Singh, Praveen Kumar CSIC ORCID; Crespo, Isidro; Gago-Córdoba, César; Hao, Jian-An; Luque-Ortega, Juan Román CSIC ORCID ; Alfonso, Carlos CSIC ORCID ; Wu, Ling J.; Boer, Roeland CSIC ORCID; Meijer, Wilfried J. J. CSIC ORCID
KeywordsAntibiotic-resistance reservoir
Protein database searches
Bacillus-subtilis
Functional-characterization
Replication origins
Plasmid PLS20
PSI-BLAST
DNA
Microbiome
Diversity
Issue Date16-Feb-2017
PublisherPublic Library of Science
CitationPLoS Genet 13 (2) e1006586 (2017)
AbstractAntibiotic resistance is a serious global problem. Antibiotic resistance genes (ARG), which are widespread in environmental bacteria, can be transferred to pathogenic bacteria via horizontal gene transfer (HGT). Gut microbiomes are especially apt for the emergence and dissemination of ARG. Conjugation is the HGT route that is predominantly responsible for the spread of ARG. Little is known about conjugative elements of Gram-positive bacteria, including those of the phylum Firmicutes, which are abundantly present in gut microbiomes. A critical step in the conjugation process is the relaxase-mediated site-and strand-specific nick in the oriT region of the conjugative element. This generates a single-stranded DNA molecule that is transferred from the donor to the recipient cell via a connecting channel. Here we identified and characterized the relaxosome components oriT and the relaxase of the conjugative plasmid pLS20 of the Firmicute Bacillus subtilis. We show that the relaxase gene, named rel(LS20), is essential for conjugation, that it can function in trans and provide evidence that Tyr26 constitutes the active site residue. In vivo and in vitro analyses revealed that the oriT is located far upstream of the relaxase gene and that the nick site within oriT is located on the template strand of the conjugation genes. Surprisingly, the Rel(LS20) shows very limited similarity to known relaxases. However, more than 800 genes to which no function had been attributed so far are predicted to encode proteins showing significant similarity to RelLS20. Interestingly, these putative relaxases are encoded almost exclusively in Firmicutes bacteria. Thus, RelLS20 constitutes the prototype of a new family of relaxases. The identification of this novel relaxase family will have an important impact in different aspects of future research in the field of HGT in Gram-positive bacteria in general, and specifically in the phylum of Firmicutes, and in gut microbiome research.
Description23 p.-5 fig.-2 tab.
Publisher version (URL)https://doi.org/10.1371/journal.pgen.1006586
URIhttp://hdl.handle.net/10261/219862
DOI10.1371/journal.pgen.1006586
ISSN1553-7390
E-ISSN1553-7404
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