ABSTRACT
T helper (Th)17 cells represent a unique subset of CD4+ T cells and are vital for clearance of extracellular pathogens including bacteria and fungi. However, Th17 cells are also involved in orchestrating autoimmunity. By employing quantitative surface proteomics, we found that the evolutionarily conserved prohibitins (PHB1/2) are highly expressed on the surface of both murine and human Th17 cells. Increased expression of PHBs at the cell surface contributed to enhanced CRAF/MAPK activation in Th17 cells. Targeting surface-expressed PHBs on Th17 cells with ligands such as Vi polysaccharide (Typhim vaccine) inhibited CRAF-MAPK pathway, reduced interleukin (IL)-17 expression and ameliorated disease pathology with an increase in FOXP3+-expressing Tregs in an animal model for multiple sclerosis (MS). Interestingly, we detected a CD4+ T cell population with high PHB1 surface expression in blood samples from MS patients in comparison with age- and sex-matched healthy subjects. Our observations suggest a pivotal role for the PHB-CRAF-MAPK signalling axis in regulating the polarization and pathogenicity of Th17 cells and unveil druggable targets in autoimmune disorders such as MS.
Subject(s)
Autoimmunity , Multiple Sclerosis/immunology , Repressor Proteins/immunology , Signal Transduction/immunology , Th17 Cells/immunology , Animals , Extracellular Signal-Regulated MAP Kinases/immunology , Forkhead Transcription Factors/immunology , HeLa Cells , Humans , Mice , Multiple Sclerosis/pathology , Prohibitins , Rickettsial Vaccines/pharmacology , T-Lymphocytes, Regulatory/immunology , T-Lymphocytes, Regulatory/pathology , Th17 Cells/pathologyABSTRACT
Mitogen-activated protein kinases (MAPKs) are highly conserved protein kinase modules, and they control fundamental cellular processes. While the activation of MAPKs has been well studied, little is known on the mechanisms driving their inactivation. Here we uncover a role for ubiquitination in the inactivation of a MAPK module. Extracellular-signal-regulated kinase 5 (ERK5) is a unique, conserved member of the MAPK family and is activated in response to various stimuli through a three-tier cascade constituting MEK5 and MEKK2/3. We reveal an unexpected role for Inhibitors of Apoptosis Proteins (IAPs) in the inactivation of ERK5 pathway in a bimodal manner involving direct interaction and ubiquitination. XIAP directly interacts with MEKK2/3 and competes with PB1 domain-mediated binding to MEK5. XIAP and cIAP1 conjugate predominantly K63-linked ubiquitin chains to MEKK2 and MEKK3 which directly impede MEK5-ERK5 interaction in a trimeric complex leading to ERK5 inactivation. Consistently, loss of XIAP or cIAP1 by various strategies leads to hyperactivation of ERK5 in normal and tumorigenic cells. Loss of XIAP promotes differentiation of human primary skeletal myoblasts to myocytes in a MEKK2/3-ERK5-dependent manner. Our results reveal a novel, obligatory role for IAPs and ubiquitination in the physical and functional disassembly of ERK5-MAPK module and human muscle cell differentiation.
Subject(s)
Inhibitor of Apoptosis Proteins/metabolism , MAP Kinase Kinase 5/metabolism , MAP Kinase Kinase Kinase 3/metabolism , MAP Kinase Kinase Kinases/metabolism , Mitogen-Activated Protein Kinase 7/metabolism , Ubiquitin/metabolism , X-Linked Inhibitor of Apoptosis Protein/metabolism , Cell Differentiation , Cells, Cultured , DNA-Binding Proteins , Enzyme Activation , Humans , Inhibitor of Apoptosis Proteins/genetics , MAP Kinase Kinase 5/genetics , MAP Kinase Kinase Kinase 2 , MAP Kinase Kinase Kinase 3/genetics , MAP Kinase Kinase Kinases/genetics , MEF2 Transcription Factors/metabolism , Mitogen-Activated Protein Kinase 7/genetics , Myoblasts/cytology , Myoblasts/metabolism , Nuclear Proteins/metabolism , Protein Multimerization , Protein Structure, Tertiary , Signal Transduction/physiology , Transcription Factors/metabolism , Ubiquitination , X-Linked Inhibitor of Apoptosis Protein/geneticsABSTRACT
Recently, we revealed that ubiquitination of MEKK2 and MEKK3 by inhibitor of apoptosis proteins (IAPs) directly disrupts MEK5/ERK5 interaction and subsequently attenuates ERK5 activation. In addition, loss of XIAP promotes human myogenic differentiation in an ERK5-dependent manner. These results reveal another layer of MAPK regulation and a novel role for XIAP in controlling myogenic differentiation.