Please use this identifier to cite or link to this item:http://hdl.handle.net/20.500.12105/7105
The structure of transcription termination factor Nrd1 reveals an original mode for GUAA recognition
Nucleic Acids Res. 2017 ;45(17):10293-10305.
Transcription termination of non-coding RNAs is regulated in yeast by a complex of three RNA binding proteins: Nrd1, Nab3 and Sen1. Nrd1 is central in this process by interacting with Rbp1 of RNA polymerase II, Trf4 of TRAMP and GUAA/G terminator sequences. We lack structural data for the last of these binding events. We determined the structures of Nrd1 RNA binding domain and its complexes with three GUAA-containing RNAs, characterized RNA binding energetics and tested rationally designed mutants in vivo. The Nrd1 structure shows an RRM domain fused with a second α/β domain that we name split domain (SD), because it is formed by two non-consecutive segments at each side of the RRM. The GUAA interacts with both domains and with a pocket of water molecules, trapped between the two stacking adenines and the SD. Comprehensive binding studies demonstrate for the first time that Nrd1 has a slight preference for GUAA over GUAG and genetic and functional studies suggest that Nrd1 RNA binding domain might play further roles in non-coding RNAs transcription termination.
Amino Acid Sequence | Conserved Sequence | Crystallography, X-Ray | Models, Molecular | Mutation | Nuclear Magnetic Resonance, Biomolecular | Peptide Fragments | Protein Binding | Protein Conformation | Protein Domains | Protein Folding | RNA, Fungal | RNA, Messenger | RNA-Binding Proteins | Recombinant Fusion Proteins | Saccharomyces cerevisiae | Saccharomyces cerevisiae Proteins | Sequence Alignment | Substrate Specificity | Transcription Termination, Genetic