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. 1997 Sep 1;16(17):5235–5246. doi: 10.1093/emboj/16.17.5235

RNA recognition by the joint action of two nucleolin RNA-binding domains: genetic analysis and structural modeling.

P Bouvet 1, C Jain 1, J G Belasco 1, F Amalric 1, M Erard 1
PMCID: PMC1170156  PMID: 9311984

Abstract

The interaction of nucleolin with a short stem-loop structure (NRE) requires two contiguous RNA-binding domains (RBD 1+2). The structural basis for RNA recognition by these RBDs was studied using a genetic system in Escherichia coli. Within each of the two domains, we identified several mutations that severely impair interaction with the RNA target. Mutations that alter RNA-binding specificity were also isolated, suggesting the identity of specific contacts between RBD 1+2 amino acids and nucleotides within the NRE stem-loop. Our data indicate that both RBDs participate in a joint interaction with the NRE and that each domain uses a different surface to contact the RNA. The constraints provided by these genetic data and previous mutational studies have enabled us to propose a three-dimensional model of nucleolin RBD 1+2 bound to the NRE stem-loop.

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