title: Kinase domain insertions define distinct roles of CLK kinases in SR protein phosphorylation.
abstract: 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.
authors.affiliation: Structural Genomics Consortium, University of Oxford, Old Road Research Building, Roosevelt Drive, Oxford OX3 7DQ, UK. alex.bullock@sgc.ox.ac.uk
authors.forename: Alex N
authors.initials: AN
authors.lastname: Bullock
authors.forename: Sanjan
authors.initials: S
authors.lastname: Das
authors.forename: Judit E
authors.initials: JE
authors.lastname: Debreczeni
authors.forename: Peter
authors.initials: P
authors.lastname: Rellos
authors.forename: Oleg
authors.initials: O
authors.lastname: Fedorov
authors.forename: Frank H
authors.initials: FH
authors.lastname: Niesen
authors.forename: Kunde
authors.initials: K
authors.lastname: Guo
authors.forename: Evangelos
authors.initials: E
authors.lastname: Papagrigoriou
authors.forename: Ann L
authors.initials: AL
authors.lastname: Amos
authors.forename: Suhyung
authors.initials: S
authors.lastname: Cho
authors.forename: Benjamin E
authors.initials: BE
authors.lastname: Turk
authors.forename: Gourisankar
authors.initials: G
authors.lastname: Ghosh
authors.forename: Stefan
authors.initials: S
authors.lastname: Knapp
mesh_terms: D000595:Amino Acid Sequence; D001665:Binding Sites; D006801:Humans; D008958:Models, Molecular; D008969:Molecular Sequence Data; D016254:Mutagenesis, Insertional; D009687:Nuclear Proteins / Q000737:chemistry / Q000378:metabolism*; D010766:Phosphorylation; D011487:Protein Conformation; D017346:Protein Serine-Threonine Kinases / Q000737:chemistry / Q000378:metabolism*; D017434:Protein Structure, Tertiary; D011505:Protein-Tyrosine Kinases / Q000737:chemistry / Q000378:metabolism*; D012326:RNA Splicing; D016601:RNA-Binding Proteins / Q000737:chemistry / Q000378:metabolism*; D000068103:Serine-Arginine Splicing Factors; D013379:Substrate Specificity
chemical_list: D009687:Nuclear Proteins; D016601:RNA-Binding Proteins; D000068103:Serine-Arginine Splicing Factors; C066780:Clk dual-specificity kinases; D011505:Protein-Tyrosine Kinases; D017346:Protein Serine-Threonine Kinases