title: Docking interactions of the JNK scaffold protein WDR62.
abstract: JNK (c-Jun N-terminal kinase) is part of a MAPK (mitogen-activated protein kinase) signalling cascade. Scaffold proteins simultaneously associate with various components of the MAPK signalling pathway and play a crucial role in signal transmission and MAPK regulation. WDR62 (WD repeat domain 62) is a JNK scaffold protein. Recessive mutations within WDR62 result in severe cerebral cortical malformation. In the present study we demonstrate the association of WDR62 with endogenous and overexpressed proteins of both JNK2 and the JNK2-activating kinase MKK7 (MAPK kinase 7). Association of WDR62 with JNK2 and MKK7 occurs via direct protein-protein interactions. We mapped the docking domain of WDR62 responsible for the association with JNK. WDR62 interacts with all JNK isoforms through a D domain motif located at the C-terminus. A WDR62 mutant lacking the putative JNK-binding domain fails to activate and recruit JNK to cellular granules. Furthermore, a synthetic peptide composed of the WDR62 docking domain inhibits JNK2 activity in vitro. WDR62 association with JNK2 requires both the JNK CD and ED domains, and the binding requisite is distinct from that of the previously described JNK2 association with JIP1 (JNK-interacting protein 1). Next, we characterized the association between WDR62 and MKK7. WDR62 associates directly with the MKK7β1 isoform independently of JNK binding, but fails to interact with MKK7α1. Furthermore, MKK7β1 recruits a protein phosphatase that dephosphorylates WDR62. Interestingly, a premature termination mutation in WDR62 that results in severe brain developmental defects does not abrogate WDR62 association with either JNK or MKK7. Therefore such mutations represent a loss of WDR62 function independent of JNK signalling.
authors.affiliation: Department of Molecular Genetics, The Rappaport Family Institute for Research in the Medical Sciences, Technion - Israel Institute of Technology, Bat-Galim, Haifa, Israel.
authors.forename: Ksenya
authors.initials: K
authors.lastname: Cohen-Katsenelson
authors.forename: Tanya
authors.initials: T
authors.lastname: Wasserman
authors.forename: Samer
authors.initials: S
authors.lastname: Khateb
authors.forename: Alan J
authors.initials: AJ
authors.lastname: Whitmarsh
authors.forename: Ami
authors.initials: A
authors.lastname: Aronheim
mesh_terms: D000595:Amino Acid Sequence; D001665:Binding Sites; D018797:Cell Cycle Proteins; D057809:HEK293 Cells; D006367:HeLa Cells; D006801:Humans; D048031:JNK Mitogen-Activated Protein Kinases / Q000737:chemistry / Q000235:genetics; D048688:MAP Kinase Kinase 7 / Q000737:chemistry* / Q000235:genetics; D048056:Mitogen-Activated Protein Kinase 9 / Q000737:chemistry* / Q000235:genetics; D008969:Molecular Sequence Data; D009419:Nerve Tissue Proteins / Q000737:chemistry* / Q000235:genetics; D034521:Nuclear Matrix-Associated Proteins / Q000737:chemistry / Q000235:genetics; D011485:Protein Binding; D017434:Protein Structure, Tertiary / Q000235:genetics; D020449:Repetitive Sequences, Amino Acid / Q000235:genetics; D017384:Sequence Deletion / Q000235:genetics
chemical_list: D018797:Cell Cycle Proteins; D009419:Nerve Tissue Proteins; D034521:Nuclear Matrix-Associated Proteins; C552527:WDR62 protein, human; D048056:Mitogen-Activated Protein Kinase 9; D048031:JNK Mitogen-Activated Protein Kinases; D048688:MAP Kinase Kinase 7; C482569:MAP2K7 protein, human