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000872604 245__ $$aDeciphering the Rules Underlying Xenogeneic Silencing and Counter-Silencing of Lsr2-like Proteins Using CgpS of Corynebacterium glutamicum as a Model
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000872604 520__ $$aLsr2-like nucleoid-associated proteins play an important role as xenogeneic silencers (XS) of horizontally acquired genomic regions in actinobacteria. In this study, we systematically analyzed the in vivo constraints underlying silencing and counter-silencing of the Lsr2-like protein CgpS in Corynebacterium glutamicum. Genome-wide analysis revealed binding of CgpS to regions featuring a distinct drop in GC profile close to the transcription start site (TSS) but also identified an overrepresented motif with multiple A/T steps at the nucleation site of the nucleoprotein complex. Binding of specific transcription factors (TFs) may oppose XS activity, leading to counter-silencing. Following a synthetic counter-silencing approach, target gene activation was realized by inserting operator sites of an effector-responsive TF within various CgpS target promoters, resulting in increased promoter activity upon TF binding. Analysis of reporter constructs revealed maximal counter-silencing when the TF operator site was inserted at the position of maximal CgpS coverage. This principle was implemented in a synthetic toggle switch, which features a robust and reversible response to effector availability, highlighting the potential for biotechnological applications. Together, our results provide comprehensive insights into how Lsr2 silencing and counter-silencing shape evolutionary network expansion in this medically and biotechnologically relevant bacterial phylum.
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000872604 7001_ $$0P:(DE-Juel1)179244$$aFilipchyk, Andrei$$b1$$ufzj
000872604 7001_ $$0P:(DE-Juel1)165889$$aHünnefeld, Max$$b2$$ufzj
000872604 7001_ $$0P:(DE-HGF)0$$aGätgens, Cornelia$$b3
000872604 7001_ $$0P:(DE-HGF)0$$aBrehm, Jannis$$b4
000872604 7001_ $$0P:(DE-HGF)0$$aHeermann, Ralf$$b5
000872604 7001_ $$0P:(DE-Juel1)138503$$aFrunzke, Julia$$b6$$eCorresponding author
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