Kinase domain insertions define distinct roles of CLK kinases in SR protein phosphorylation
Splicing requires reversible phosphorylation of serine/arginine-rich (SR) proteins, which direct splice site selection in eukaryotic mRNA. These phosphorylation events are dependent on SR protein (SRPK) and cdc2-like kinase (CLK) families. SRPK1 phosphorylation of splicing factors is restricted by a...
| Main Authors: | , , , , , , , , , , , , |
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| Format: | Journal article |
| Language: | English |
| Published: |
Cell Press
2009
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| Subjects: |
| _version_ | 1797054502389415936 |
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| author | Bullock, A Das, S Debreczeni, J Rellos, P Fedorov, O Niesen, F Guo, K Papagrigoriou, E Amos, A Cho, S Turk, B Ghosh, G Knapp, S |
| author_facet | Bullock, A Das, S Debreczeni, J Rellos, P Fedorov, O Niesen, F Guo, K Papagrigoriou, E Amos, A Cho, S Turk, B Ghosh, G Knapp, S |
| author_sort | Bullock, A |
| collection | OXFORD |
| description | Splicing requires reversible phosphorylation of serine/arginine-rich (SR) proteins, which direct splice site selection in eukaryotic mRNA. These phosphorylation events are dependent on SR protein (SRPK) and cdc2-like kinase (CLK) families. SRPK1 phosphorylation of splicing factors is restricted by a specific docking interaction whereas CLK activity is less constrained. To understand functional differences between splicing factor targeting kinases, we determined crystal structures of CLK1 and CLK3. Intriguingly, in CLKs the SRPK1 docking site is blocked by insertion of a previously unseen helix alphaH. In addition, substrate docking grooves present in related mitogen activating protein kinases (MAPKs) are inaccessible due to a CLK specific beta7/8-hairpin insert. Thus, the unconstrained substrate interaction together with the determined active-site mediated substrate specificity allows CLKs to complete the functionally important hyperphosphorylation of splicing factors like ASF/SF2. In addition, despite high sequence conservation, we identified inhibitors with surprising isoform specificity for CLK1 over CLK3. |
| first_indexed | 2024-03-06T18:58:05Z |
| format | Journal article |
| id | oxford-uuid:128af0bf-7e01-4188-b305-6295064a2571 |
| institution | University of Oxford |
| language | English |
| last_indexed | 2024-03-06T18:58:05Z |
| publishDate | 2009 |
| publisher | Cell Press |
| record_format | dspace |
| spelling | oxford-uuid:128af0bf-7e01-4188-b305-6295064a25712022-03-26T10:08:38ZKinase domain insertions define distinct roles of CLK kinases in SR protein phosphorylationJournal articlehttp://purl.org/coar/resource_type/c_dcae04bcuuid:128af0bf-7e01-4188-b305-6295064a2571PhosphorylationRNA-Binding ProteinsProtein-Tyrosine KinasesRNA SplicingHumansProtein-Serine-Threonine KinasesProtein Structure, TertiarymetabolismMutagenesis, InsertionalProtein ConformationNuclear ProteinsMolecular Sequence DataBinding SitesModels, MolecularAmino Acid SequencechemistrySubstrate SpecificityEnglishStructural Genomics ConsortiumCell Press2009Bullock, ADas, SDebreczeni, JRellos, PFedorov, ONiesen, FGuo, KPapagrigoriou, EAmos, ACho, STurk, BGhosh, GKnapp, SSplicing requires reversible phosphorylation of serine/arginine-rich (SR) proteins, which direct splice site selection in eukaryotic mRNA. These phosphorylation events are dependent on SR protein (SRPK) and cdc2-like kinase (CLK) families. SRPK1 phosphorylation of splicing factors is restricted by a specific docking interaction whereas CLK activity is less constrained. To understand functional differences between splicing factor targeting kinases, we determined crystal structures of CLK1 and CLK3. Intriguingly, in CLKs the SRPK1 docking site is blocked by insertion of a previously unseen helix alphaH. In addition, substrate docking grooves present in related mitogen activating protein kinases (MAPKs) are inaccessible due to a CLK specific beta7/8-hairpin insert. Thus, the unconstrained substrate interaction together with the determined active-site mediated substrate specificity allows CLKs to complete the functionally important hyperphosphorylation of splicing factors like ASF/SF2. In addition, despite high sequence conservation, we identified inhibitors with surprising isoform specificity for CLK1 over CLK3. |
| spellingShingle | Phosphorylation RNA-Binding Proteins Protein-Tyrosine Kinases RNA Splicing Humans Protein-Serine-Threonine Kinases Protein Structure, Tertiary metabolism Mutagenesis, Insertional Protein Conformation Nuclear Proteins Molecular Sequence Data Binding Sites Models, Molecular Amino Acid Sequence chemistry Substrate Specificity Bullock, A Das, S Debreczeni, J Rellos, P Fedorov, O Niesen, F Guo, K Papagrigoriou, E Amos, A Cho, S Turk, B Ghosh, G Knapp, S Kinase domain insertions define distinct roles of CLK kinases in SR protein phosphorylation |
| title | Kinase domain insertions define distinct roles of CLK kinases in SR protein phosphorylation |
| title_full | Kinase domain insertions define distinct roles of CLK kinases in SR protein phosphorylation |
| title_fullStr | Kinase domain insertions define distinct roles of CLK kinases in SR protein phosphorylation |
| title_full_unstemmed | Kinase domain insertions define distinct roles of CLK kinases in SR protein phosphorylation |
| title_short | Kinase domain insertions define distinct roles of CLK kinases in SR protein phosphorylation |
| title_sort | kinase domain insertions define distinct roles of clk kinases in sr protein phosphorylation |
| topic | Phosphorylation RNA-Binding Proteins Protein-Tyrosine Kinases RNA Splicing Humans Protein-Serine-Threonine Kinases Protein Structure, Tertiary metabolism Mutagenesis, Insertional Protein Conformation Nuclear Proteins Molecular Sequence Data Binding Sites Models, Molecular Amino Acid Sequence chemistry Substrate Specificity |
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