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Δευτέρα 18 Ιουνίου 2018

Genes and Proteins Involved in qnrS1 Induction [PublishAheadOfPrint]

Expression of the quinolone resistance gene qnrS1 is increased by quinolones, but, unlike induction of some other qnr genes, the bacterial SOS system is not involved and no lexA box is found upstream. Nonetheless, at least 205-bp of upstream sequence is required for induction to take place. An upstream sequence bound to beads trapped potential binding proteins from cell extracts that were identified by mass spectrometry as Dps, Fis, Ihf, Lrp, CysB, and YjhU. To further elucidate their role a reporter plasmid linking the qnrS1 upstream sequence to lacZ was introduced into cells of the Keio collection with single gene deletions and screened for lacZ expression. Mutants in ihfA and ihfB had decreased lacZ induction while induction in a cysB mutant was increased and dps, fis, lrp, yjhU and other mutants showed no change. The essential upstream sequence contains potential binding sites for Ihf and DnaA. A dnaA deletion could not be tested because it provides essential functions in cell replication, but increased dnaA expression decreased qnrS1 induction while decreased dnaA expression enhanced it implying a role for DnaA as a repressor. In a mobility shift assay purified IhfA, IhfB, and DnaA proteins (but not CysB) were shown to bind to the upstream segment. Induction decreased in a gyrA quinolone-resistant mutant indicating that GyrA also has a role. Thus, quinolones acting through proteins DnaA, GyrA, IhfA, and IhfB regulates expression of qnrS1.



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