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.

journal_name

RNA Biol

journal_title

RNA biology

authors

Nozinovic S,Reining A,Kim YB,Noeske J,Schlepckow K,Wöhnert J,Schwalbe H

doi

10.4161/rna.29439

subject

Has Abstract

pub_date

2014-01-01 00:00:00

pages

655-6

issue

5

eissn

1547-6286

issn

1555-8584

pii

29439

journal_volume

11

pub_type

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