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Engineering RNA sequence specificity of Pumilio repeats.


ABSTRACT: Puf proteins bind RNA sequence specifically and regulate translation and stability of target mRNAs. A "code" for RNA recognition has been deduced from crystal structures of the Puf protein, human Pumilio1, where each of eight repeats binds an RNA base via a combination of three side chains at conserved positions. Here, we report the creation of seven soluble mutant proteins with predictably altered sequence specificity, including one that binds tightly to adenosine-uracil-rich element RNA. These data show that Pumilio1 can be used as a scaffold to engineer RNA-binding proteins with designed sequence specificity.

SUBMITTER: Cheong CG 

PROVIDER: S-EPMC1564246 | biostudies-literature | 2006 Sep

REPOSITORIES: biostudies-literature

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Engineering RNA sequence specificity of Pumilio repeats.

Cheong Cheom-Gil CG   Hall Traci M Tanaka TM  

Proceedings of the National Academy of Sciences of the United States of America 20060905 37


Puf proteins bind RNA sequence specifically and regulate translation and stability of target mRNAs. A "code" for RNA recognition has been deduced from crystal structures of the Puf protein, human Pumilio1, where each of eight repeats binds an RNA base via a combination of three side chains at conserved positions. Here, we report the creation of seven soluble mutant proteins with predictably altered sequence specificity, including one that binds tightly to adenosine-uracil-rich element RNA. These  ...[more]

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