Specific and modular binding code for cytosine recognition in Pumilio/FBF (PUF) RNA-binding domains.

ABSTRACT: Pumilio/fem-3 mRNA-binding factor (PUF) proteins possess a recognition code for bases A, U, and G, allowing designed RNA sequence specificity of their modular Pumilio (PUM) repeats. However, recognition side chains in a PUM repeat for cytosine are unknown. Here we report identification of a cytosine-recognition code by screening random amino acid combinations at conserved RNA recognition positions using a yeast three-hybrid system. This C-recognition code is specific and modular as specificity can be transferred to different positions in the RNA recognition sequence. A crystal structure of a modified PUF domain reveals specific contacts between an arginine side chain and the cytosine base. We applied the C-recognition code to design PUF domains that recognize targets with multiple cytosine

SUBMITTER: Dong S 

PROVIDER: S-EPMC3144504 | biostudies-literature | 2011 Jul

REPOSITORIES: biostudies-literature