SF1 Phosphorylation Enhances Specific Binding to U2AF⁶⁵ and Reduces Binding to 3'-Splice-Site RNA.

ABSTRACT: Splicing factor 1 (SF1) recognizes 3' splice sites of the major class of introns as a ternary complex with U2AF⁶⁵ and U2AF³⁵ splicing factors. A conserved SPSP motif in a coiled-coil domain of SF1 is highly phosphorylated in proliferating human cells and is required for cell proliferation. The UHM kinase 1 (UHMK1), also called KIS, double-phosphorylates both serines of this SF1 motif. Here, we use isothermal titration calorimetry to demonstrate that UHMK1 phosphorylation of the SF1 SPSP motif slightly enhances specific binding of phospho-SF1 to its cognate U2AF⁶⁵ protein partner. Conversely, quantitative fluorescence anisotropy RNA binding assays and isothermal titration calorimetry experiments establish that double-SPSP phosphorylation reduces phospho-SF1

SUBMITTER: Chatrikhi R 

PROVIDER: S-EPMC5192697 | biostudies-literature | 2016 Dec

REPOSITORIES: biostudies-literature