Medizinische Universität Graz Austria/Österreich - Forschungsportal - Medical University of Graz
Gewählte Publikation:
SHR Neuro Krebs Kardio Lipid Stoffw Microb
Zhang, Y; Madl, T; Bagdiul, I; Kern, T; Kang, HS; Zou, P; Mäusbacher, N; Sieber, SA; Krämer, 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. 2013; 41(2): 1343-1354.
Doi: 10.1093/nar/gks1097
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- Co-Autor*innen der Med Uni Graz
- Madl Tobias
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- 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.
- Find related publications in this database (using NLM MeSH Indexing)
- Amino Acid Sequence -
- DNA-Binding Proteins - chemistry DNA-Binding Proteins - metabolism
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- Models, Molecular -
- Molecular Sequence Data -
- Nuclear Proteins - chemistry Nuclear Proteins - metabolism
- Phosphorylation -
- Protein Binding -
- Protein Interaction Domains and Motifs -
- Protein Structure, Secondary -
- RNA - metabolism
- RNA Splice Sites -
- Ribonucleoproteins - chemistry Ribonucleoproteins - metabolism
- Sequence Homology, Amino Acid -
- Serine - metabolism
- Transcription Factors - chemistry Transcription Factors - metabolism