An extended U2AF65–RNA-binding domain recognizes the 3′ splice site signal
| Autor: | Enea Salsi, Anant A. Agrawal, Jermaine L. Jenkins, Clara L. Kielkopf, Rakesh Chatrikhi, Steven Henderson, Dmitri N. Ermolenko, Michael R. Green |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2016 |
| Předmět: |
0301 basic medicine
RNA Splicing Science General Physics and Astronomy Computational biology Biology Crystallography X-Ray General Biochemistry Genetics and Molecular Biology Article 03 medical and health sciences SR protein Splicing Factor U2AF RNA Precursors Humans Genetics Multidisciplinary Splice site mutation Intron Nuclear Proteins General Chemistry Protein Structure Tertiary 030104 developmental biology Polypyrimidine tract Ribonucleoproteins RNA editing RNA splicing RNA Splice Sites Small nuclear RNA |
| Zdroj: | Nature Communications, Vol 7, Iss 1, Pp 1-14 (2016) Nature Communications |
| ISSN: | 2041-1723 |
| Popis: | How the essential pre-mRNA splicing factor U2AF65 recognizes the polypyrimidine (Py) signals of the major class of 3′ splice sites in human gene transcripts remains incompletely understood. We determined four structures of an extended U2AF65–RNA-binding domain bound to Py-tract oligonucleotides at resolutions between 2.0 and 1.5 Å. These structures together with RNA binding and splicing assays reveal unforeseen roles for U2AF65 inter-domain residues in recognizing a contiguous, nine-nucleotide Py tract. The U2AF65 linker residues between the dual RNA recognition motifs (RRMs) recognize the central nucleotide, whereas the N- and C-terminal RRM extensions recognize the 3′ terminus and third nucleotide. Single-molecule FRET experiments suggest that conformational selection and induced fit of the U2AF65 RRMs are complementary mechanisms for Py-tract association. Altogether, these results advance the mechanistic understanding of molecular recognition for a major class of splice site signals. The pre-mRNA splicing factor U2AF65 recognizes 3′ splice sites in human gene transcripts, but the details are not fully understood. Here, the authors report U2AF65 structures and single molecule FRET that reveal mechanistic insights into splice site recognition. |
| Databáze: | OpenAIRE |
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