Thus, this finding highlights a global conformational change associated with substrate binding in the VS ribozyme that is likely critical for its enzymatic activity. However, a local structural difference is observed that affects the global fold by shifting the relative orientation of the two three-way junctions. The resulting NMR-SAXS structural ensemble shares several similarities with the reported crystal structures of the VS ribozyme. These subdomain structures were then assembled to build a large set of structural models of the ribozyme, which was subsequently filtered using SAXS data. A multi-step procedure was used for structure determination that first involved pairing refined NMR structures with SAXS data to obtain structural subensembles of the various subdomains. NMR and SAXS studies were conducted on a minimal trans VS ribozyme as well as several isolated subdomains. Here, we introduce an integrative approach based on the divide-and-conquer strategy that was undertaken to determine the solution structure of an RNA model system, the Neurospora VS ribozyme. The divide-and-conquer strategy is commonly used for protein structure determination, but its applications to high-resolution structure determination of RNAs have been limited.
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