5EV3

Structure III of Intact U2AF65 Recognizing the 3' Splice Site Signal

Summary for 5EV3

• Entry DOI: 10.2210/pdb5ev3/pdb
• Related: 5EV1 5EV2 5EV4
• Descriptor: Splicing factor U2AF 65 kDa subunit, DNA/RNA (5'-R(P*UP*U)-D(P*U)-R(P*UP*U)-D(P*(BRU)P*UP*U)-3') (3 entities in total)
• Functional Keywords: protein-rna complex, rna splicing factor, rna recognition motif, polypyrimidine tract, rna binding protein-rna complex, rna binding protein/rna
• Biological source: Homo sapiens (Human); More
• Total number of polymer chains: 2
• Total formula weight: 24389.27

Authors

Agrawal, A.A.,Jenkins, J.L.,Kielkopf, C.L. (deposition date: 2015-11-19, release date: 2016-02-24, Last modification date: 2024-03-06)

Primary citation

Agrawal, A.A.,Salsi, E.,Chatrikhi, R.,Henderson, S.,Jenkins, J.L.,Green, M.R.,Ermolenko, D.N.,Kielkopf, C.L.
An extended U2AF(65)-RNA-binding domain recognizes the 3' splice site signal.
Nat Commun, 7:10950-10950, 2016

PubMed Abstract: How the essential pre-mRNA splicing factor U2AF(65) 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 U2AF(65)-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 U2AF(65) inter-domain residues in recognizing a contiguous, nine-nucleotide Py tract. The U2AF(65) 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 U2AF(65) 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.

PubMed: 26952537
DOI: 10.1038/ncomms10950

Experimental method

X-RAY DIFFRACTION (1.5 Å)