2M0G

Structure, phosphorylation and U2AF65 binding of the Nterminal Domain of splicing factor 1 during 3 splice site Recognition

Summary for 2M0G

• Entry DOI: 10.2210/pdb2m0g/pdb
• NMR Information: BMRB: 18808
• Descriptor: Splicing factor 1, Splicing factor U2AF 65 kDa subunit (2 entities in total)
• Functional Keywords: spliceosome assembly, sf1, uhm, ulm, splicing
• Biological source: Homo sapiens (human); More
• Total number of polymer chains: 2
• Total formula weight: 28651.68

Authors

Madl, T.,Sattler, M.,Zhang, Y.,Bagdiul, I.,Kern, T.,Kang, H.,Zou, P.,Maeusbacher, N.,Sieber, S.A.,Kraemer, A. (deposition date: 2012-10-25, release date: 2013-01-30, Last modification date: 2024-05-01)

Primary citation

Zhang, Y.,Madl, T.,Bagdiul, I.,Kern, T.,Kang, H.S.,Zou, P.,Mausbacher, N.,Sieber, S.A.,Kramer, A.,Sattler, M.
Structure, phosphorylation and U2AF65 binding of the N-terminal domain of splicing factor 1 during 3'-splice site recognition.
Nucleic Acids Res., 41:1343-1354, 2013

PubMed Abstract: Recognition of the 3'-splice site is a key step in pre-mRNA splicing and accomplished by a dynamic complex comprising splicing factor 1 (SF1) and the U2 snRNP auxiliary factor 65-kDa subunit (U2AF65). Both proteins mediate protein-protein and protein-RNA interactions for cooperative RNA-binding during spliceosome assembly. Here, we report the solution structure of a novel helix-hairpin domain in the N-terminal region of SF1 (SF1(NTD)). The nuclear magnetic resonance- and small-angle X-ray scattering-derived structure of a complex of the SF1(NTD) with the C-terminal U2AF homology motif domain of U2AF65 (U2AF65(UHM)) reveals that, in addition to the known U2AF65(UHM)-SF1 interaction, the helix-hairpin domain forms a secondary, hydrophobic interface with U2AF65(UHM), which locks the orientation of the two subunits. Mutational analysis shows that the helix hairpin is essential for cooperative formation of the ternary SF1-U2AF65-RNA complex. We further show that tandem serine phosphorylation of a conserved Ser80-Pro81-Ser82-Pro83 motif rigidifies a long unstructured linker in the SF1 helix hairpin. Phosphorylation does not significantly alter the overall conformations of SF1, SF1-U2AF65 or the SF1-U2AF65-RNA complexes, but slightly enhances RNA binding. Our results indicate that the helix-hairpin domain of SF1 is required for cooperative 3'-splice site recognition presumably by stabilizing a unique quaternary arrangement of the SF1-U2AF65-RNA complex.

PubMed: 23175611
DOI: 10.1093/nar/gks1097

Experimental method

SOLUTION NMR