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Nick Reiter
Last active: 1/21/2015

Structure and interactions of the first three RNA recognition motifs of splicing factor prp24.

Bae E, Reiter NJ, Bingman CA, Kwan SS, Lee D, Phillips GN, Butcher SE, Brow DA
J Mol Biol. 2007 367 (5): 1447-58

PMID: 17320109 · PMCID: PMC1939982 · DOI:10.1016/j.jmb.2007.01.078

The essential Saccharomyces cerevisiae pre-messenger RNA splicing protein 24 (Prp24) has four RNA recognition motifs (RRMs) and facilitates U6 RNA base-pairing with U4 RNA during spliceosome assembly. Prp24 is a component of the free U6 small nuclear ribonucleoprotein particle (snRNP) but not the U4/U6 bi-snRNP, and so is thought to be displaced from U6 by U4/U6 base-pairing. The interaction partners of each of the four RRMs of Prp24 and how these interactions direct U4/U6 pairing are not known. Here we report the crystal structure of the first three RRMs and the solution structure of the first two RRMs of Prp24. Strikingly, RRM 2 forms extensive inter-domain contacts with RRMs 1 and 3. These contacts occupy much of the canonical RNA-binding faces (beta-sheets) of RRMs 1 and 2, but leave the beta-sheet of RRM 3 exposed. Previously identified substitutions in Prp24 that suppress mutations in U4 and U6 spliceosomal RNAs cluster primarily in the beta-sheet of RRM 3, but also in a conserved loop of RRM 2. RNA binding assays and chemical shift mapping indicate that a large basic patch evident on the surface of RRMs 1 and 2 is part of a high affinity U6 RNA binding site. Our results suggest that Prp24 binds free U6 RNA primarily with RRMs 1 and 2, which may remodel the U6 secondary structure. The beta-sheet of RRM 3 then influences U4/U6 pairing through interaction with an unidentified ligand.

MeSH Terms (16)

Amino Acid Motifs Amino Acid Sequence Models, Molecular Molecular Sequence Data Mutation Nucleic Acid Conformation Protein Binding Protein Structure, Secondary Ribonucleoproteins, Small Nuclear RNA, Fungal RNA, Small Nuclear RNA-Binding Proteins RNA Splicing Saccharomyces cerevisiae Saccharomyces cerevisiae Proteins Sequence Homology, Amino Acid

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