A scanning peptide array approach uncovers association sites within the JNK/βarrestin signalling complex

Abstract βarrestins are molecular scaffolds that can bring together three-component mitogen-activated protein kinase signalling modules to promote signal compartmentalisation. We use peptide array technology to define novel interfaces between components within the c-Jun N-terminal kinase (JNK)/βarrestin signalling complex. We show that βarrestin 1 and βarrestin 2 associate with JNK3 via the kinase N-terminal domain in a region that, surprisingly, does not harbour a known ‘common docking’ motif. In the N-domain and C-terminus of βarrestin 1 and βarrestin 2 we identify two novel apoptosis signal-regulating kinase 1 binding sites and in the N-domain of the βarrestin 1 and βarrestin 2 we identify a novel MKK4 docking site. Structured summary MINT- 7263196 , MINT- 7263175 : Arrestin beta-2 (uniprotkb: P32121 ) binds (MI: 0407 ) to ASK1 (uniprotkb: Q99683 ) by peptide array (MI: 0081 ) MINT- 7263136 : JNK3 (uniprotkb: P53779 ) binds (MI: 0407 ) to Arrestin beta-1 (uniprotkb: P49407 ) by peptide array (MI: 0081 ) MINT- 7263161 : JNK3 (uniprotkb: P53779 ) binds (MI: 0407 ) to Arrestin beta-2 (uniprotkb: P32121 ) by peptide array (MI: 0081 ) MINT- 7263304 : Arrestin beta-1 (uniprotkb: P49407 ) physically interacts (MI: 0915 ) with ASK1 (uniprotkb: Q99683 ) by anti tag coimmunoprecipitation (MI: 0007 ) MINT- 7263286 : Arrestin beta-2 (uniprotkb: P32121 ) binds (MI: 0407 ) to MKK4 (uniprotkb: P45985 ) by peptide array (MI: 0081 ) MINT- 7263231 , MINT- 7263254 : Arrestin beta-1 (uniprotkb: P49407 ) binds (MI: 0407 ) to ASK1 (uniprotkb: Q99683 ) by peptide array (MI: 0081 ) MINT- 7263269 : Arrestin beta-1 (uniprotkb: P49407 ) binds (MI: 0407 ) to MKK4 (uniprotkb: P45985 ) by peptide array (MI: 0081 )