title: Regulating SR protein phosphorylation through regions outside the kinase domain of SRPK1.
abstract: SR proteins (splicing factors containing arginine-serine repeats) are essential splicing factors whose phosphorylation by the SR-specific protein kinase (SRPK) family regulates nuclear localization and mRNA processing activity. In addition to an N-terminal extension with unknown function, SRPKs contain a large, nonhomologous spacer insert domain (SID) that bifurcates the kinase domain and anchors the kinase in the cytoplasm through interactions with chaperones. While structures for the kinase domain are now available, constructs that include regions outside this domain have been resistant to crystallographic elucidation. To investigate the conformation of the full-length kinase and the functional role of noncatalytic regions, we performed hydrogen-deuterium exchange and steady-state kinetic experiments on SRPK1. Unlike the kinase core, the large SID lacks stable, hydrogen-bonded structure and may provide an intrinsically disordered region for chaperone interactions. Conversely, the N-terminus, which positively regulates SR protein binding, adopts a stable structure when the insert domain is present and stabilizes a docking groove in the large lobe of the kinase domain. The N-terminus and SID equally enhance SR protein turnover by altering the stability of several catalytic loop segments. These studies reveal that SRPK1 uses an N-terminal extension and a large, intrinsically disordered region juxtaposed to a stable structure to facilitate high-affinity SR protein interactions and phosphorylation rates.
authors: Department of Pharmacology, University of California, San Diego, La Jolla, CA 92093, USA.
authors: Ryan M
authors: RM
authors: Plocinik
authors: Sheng
authors: S
authors: Li
authors: Tong
authors: T
authors: Liu
authors: Kendra L
authors: KL
authors: Hailey
authors: Jennifer
authors: J
authors: Whitesides
authors: Jennifer
authors: J
authors: Whitehouse
authors: Chen-Ting
authors: CT
authors: Ma
authors: Xiang-Dong
authors: XD
authors: Fu
authors: Gourisankar
authors: G
authors: Gosh
authors: Virgil L
authors: VL
authors: Woods
authors: Patricia A
authors: PA
authors: Jennings
authors: Joseph A
authors: JA
authors: Adams
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