title: TrhR, TrhY and HtdA, a novel regulatory circuit that modulates conjugation of the IncHI plasmids.
abstract: Bacterial conjugation promotes horizontal gene transfer and, consequently, the acquisition of new capabilities such as resistance to antimicrobial compounds and virulence related traits. Conjugative plasmids belonging to the incompatibility group HI are associated with multidrug resistance in Gram-negative pathogens. IncHI plasmid conjugation is thermodependent and all transfer-related genes are encoded in six operons (tra operons). Using R27, the prototype of IncHI1 plasmids, we reported that the plasmid-encoded factor HtdA represses four of the six tra operons. Moreover, our results indicated that other R27 factors were required for appropriate expression of the tra genes. In this report, using R27 libraries and random mutagenesis assays, two genes - trhR and trhY - have been identified as essential for the transcriptional expression of four tra operons and, accordingly, for the R27 conjugation. TrhR and TrhY are required simultaneously and their stimulatory activity is counteracted by HtdA. Functional and physical interactions between TrhR, TrhY and HtdA suggest that they form a three-element regulatory circuit that controls conjugation of IncHI plasmids. Expression studies suggest that H-NS represses conjugation at high temperature by repressing trhR expression. Remarkably, we show that this regulatory circuit is highly conserved among the IncHI plasmids.
authors.affiliation: Departament de Microbiologia, Universitat de Barcelona, Avda. Diagonal 643, 08028, Barcelona, Spain.
authors.forename: M
authors.initials: M
authors.lastname: Gibert
authors.forename: A
authors.initials: A
authors.lastname: Juárez
authors.forename: E L
authors.initials: EL
authors.lastname: Zechner
authors.forename: C
authors.initials: C
authors.lastname: Madrid
authors.forename: C
authors.initials: C
authors.lastname: Balsalobre