NMR resonance assignments of an engineered neomycin-sensing riboswitch RNA bound to ribostamycin and tobramycin
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The neomycin-sensing riboswitch is an engineered riboswitch developed to regulate gene expression in vivo in the lower eukaryote Saccharomyces cerevisiae upon binding to neomycin B. With a size of only 27nt it is the smallest functional riboswitch element identified so far. It binds not only neomycin B but also related aminoglycosides of the 2′-deoxystreptamine class with high affinity. The regulatory activity, however, strongly depends on the identity of the aminoglycoside. As a prerequisite for the structure determination of riboswitch-ligand complexes we report here the 1H, 15N, 13C and partial 31P chemical shift assignments for the minimal functional 27nt neomycin sensing riboswitch RNA in complex with the 4,5-linked neomycin analog ribostamycin and the 4,6-linked aminoglycoside tobramycin.
KeywordsRNA NMR-assignments Triple resonance experiments Riboswitch Aminoglycosides Gene expression/regulation
We thank Sabine Häfner and Matthias Görlach for a generous gift of T7-RNA-polymerase and Christian Richter for excellent technical support. We are grateful to the Center of Biomolecular Magnetic Resonance (BMRZ) of the Johann-Wolfgang-Goethe-University Frankfurt and the Deutsche Forschungsgemeinschaft (DFG) (grants WO 901/1-2 to J. W. and SU 402/4-1 to B.S.) for financial support. J. W. and B. S. are recipients of Aventis Foundation endowed professorships in Chemical Biology.
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