title: The importance of helix P1 stability for structural pre-organization and ligand binding affinity of the adenine riboswitch aptamer domain.
abstract: We report here an in-depth characterization of the aptamer domain of the transcriptional adenine-sensing riboswitch (pbuE) by NMR and fluorescence spectroscopy. By NMR studies, the structure of two aptamer sequences with different lengths of the helix P1, the central element involved in riboswitch conformational switching, was characterized. Hydrogen-bond interactions could be mapped at nucleotide resolution providing information about secondary and tertiary structure, structure homogeneity and dynamics. Our study reveals that the elongation of helix P1 has pronounced effects not only on the local but on the global structure of the apo aptamer domain. The structural differences induced by stabilizing helix P1 were found to be linked to changes of the ligand binding affinity as revealed from analysis of kinetic and thermodynamic data obtained from stopped-flow fluorescence studies. The results provide new insight into the sequence-dependent fine tuning of the structure and function of purine-sensing riboswitches.
authors.affiliation: Johann Wolfgang Goethe-University Frankfurt; Center for Biomolecular Magnetic Resonance (BMRZ); Max-von-Laue-Strasse 7; Frankfurt, Germany; Institute of Organic Chemistry and Chemical Biology.
authors.forename: Senada
authors.initials: S
authors.lastname: Nozinovic
authors.affiliation: Johann Wolfgang Goethe-University Frankfurt; Center for Biomolecular Magnetic Resonance (BMRZ); Max-von-Laue-Strasse 7; Frankfurt, Germany; Institute of Organic Chemistry and Chemical Biology.
authors.forename: Anke
authors.initials: A
authors.lastname: Reining
authors.affiliation: Johann Wolfgang Goethe-University Frankfurt; Center for Biomolecular Magnetic Resonance (BMRZ); Max-von-Laue-Strasse 7; Frankfurt, Germany; Institute of Organic Chemistry and Chemical Biology.
authors.forename: Yong-Boum
authors.initials: YB
authors.lastname: Kim
authors.affiliation: Department of Molecular and Cell Biology; University of California at Berkeley; Berkeley, CA USA.
authors.forename: Jonas
authors.initials: J
authors.lastname: Noeske
authors.affiliation: Johann Wolfgang Goethe-University Frankfurt; Center for Biomolecular Magnetic Resonance (BMRZ); Max-von-Laue-Strasse 7; Frankfurt, Germany; Institute of Organic Chemistry and Chemical Biology.
authors.forename: Kai
authors.initials: K
authors.lastname: Schlepckow
authors.affiliation: Johann Wolfgang Goethe-University Frankfurt; Center for Biomolecular Magnetic Resonance (BMRZ); Max-von-Laue-Strasse 7; Frankfurt, Germany; Institute for Molecular Biosciences; Max-von-Laue-Strasse 9; Frankfurt, Germany.
authors.forename: Jens
authors.initials: J
authors.lastname: Wöhnert
authors.affiliation: Johann Wolfgang Goethe-University Frankfurt; Center for Biomolecular Magnetic Resonance (BMRZ); Max-von-Laue-Strasse 7; Frankfurt, Germany; Institute of Organic Chemistry and Chemical Biology.
authors.forename: Harald
authors.initials: H
authors.lastname: Schwalbe
mesh_terms: D000225:Adenine / Q000378:metabolism*; D052157:Aptamers, Nucleotide / Q000378:metabolism*; D020029:Base Pairing; D008024:Ligands; D008274:Magnesium / Q000378:metabolism; D008958:Models, Molecular; D019906:Nuclear Magnetic Resonance, Biomolecular; D009690:Nucleic Acid Conformation; D020871:RNA Stability; D012329:RNA, Bacterial / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D058928:Riboswitch; D013816:Thermodynamics
chemical_list: D052157:Aptamers, Nucleotide; D008024:Ligands; D012329:RNA, Bacterial; D058928:Riboswitch; D008274:Magnesium; D000225:Adenine