Expanding the binding specificity for RNA recognition by a PUF domain View Full Text


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Article Info

DATE

2021-08-24

AUTHORS

Wei Zhou, Daniel Melamed, Gabor Banyai, Cindy Meyer, Thomas Tuschl, Marvin Wickens, Junyue Cao, Stanley Fields

ABSTRACT

The ability to design a protein to bind specifically to a target RNA enables numerous applications, with the modular architecture of the PUF domain lending itself to new RNA-binding specificities. For each repeat of the Pumilio-1 PUF domain, we generate a library that contains the 8,000 possible combinations of amino acid substitutions at residues critical for RNA contact. We carry out yeast three-hybrid selections with each library against the RNA recognition sequence for Pumilio-1, with any possible base present at the position recognized by the randomized repeat. We use sequencing to score the binding of each variant, identifying many variants with highly repeat-specific interactions. From these data, we generate an RNA binding code specific to each repeat and base. We use this code to design PUF domains against 16 RNAs, and find that some of these domains recognize RNAs with two, three or four changes from the wild type sequence. More... »

PAGES

5107

References to SciGraph publications

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  • Identifiers

    URI

    http://scigraph.springernature.com/pub.10.1038/s41467-021-25433-6

    DOI

    http://dx.doi.org/10.1038/s41467-021-25433-6

    DIMENSIONS

    https://app.dimensions.ai/details/publication/pub.1140616244

    PUBMED

    https://www.ncbi.nlm.nih.gov/pubmed/34429425


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