title: Protein and RNA dynamics play key roles in determining the specific recognition of GU-rich polyadenylation regulatory elements by human Cstf-64 protein.
abstract: The N-terminal domain of the 64 kDa subunit of the cleavage stimulation factor (CstF-64) recognizes GU-rich elements within the 3'-untranslated region of eukaryotic mRNAs. This interaction is essential for mRNA 3' end processing and transcription termination, and its strength affects the efficiency of utilization of different polyadenylation sites. The structure of the RNA-binding N-terminal domain of CstF-64 showed how the N-terminal RNA recognition motif of CstF-64 recognizes GU-rich RNAs. However, it is still perplexing how this protein can bind selectively to RNAs that are rich in G and U residues regardless of their detailed sequence composition, yet discriminate effectively against non-GU-RNAs. We investigated by NMR the dynamics of the CstF-64 RNA-binding domain, both free and bound to two GU-rich RNA sequences that represent polyadenylation regulatory elements. While the free protein displays the motional properties typical of a well-folded protein domain and is uniformly rigid, the protein-RNA interface acquires significant mobility on the micro- to millisecond time-scale once GU-rich RNAs binds to it. These motional features, we propose, are intrinsic to the functional requirement to bind all GU-rich sequences and yet to discriminate against non-GU-rich RNAs. This behavior may be a general mechanism by which some RNA-binding proteins are able to bind to classes of sequences, as opposed to a well-defined sequence or consensus.
authors.affiliation: Department of Biochemistry and Department of Chemistry, University of Washington, Seattle, WA 98195-1700, USA.
authors.forename: Pritilekha
authors.initials: P
authors.lastname: Deka
authors.forename: P K
authors.initials: PK
authors.lastname: Rajan
authors.forename: Jose Manuel
authors.initials: JM
authors.lastname: Perez-Canadillas
authors.forename: Gabriele
authors.initials: G
authors.lastname: Varani
mesh_terms: D039241:Cleavage Stimulation Factor; D006151:Guanosine / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D008958:Models, Molecular; D009038:Motion; D019906:Nuclear Magnetic Resonance, Biomolecular; D026723:Polyadenylation / Q000235:genetics*; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D012313:RNA / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D016601:RNA-Binding Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D013379:Substrate Specificity; D014529:Uridine / Q000235:genetics / Q000378:metabolism*
chemical_list: C474466:CSTF2T protein, human; D039241:Cleavage Stimulation Factor; D016601:RNA-Binding Proteins; D006151:Guanosine; D012313:RNA; D014529:Uridine