title: Structural requirements for recognition of major histocompatibility complex class II by membrane-associated RING-CH (MARCH) protein E3 ligases.
abstract: MARCH E3 ligases play a key role in controlling MHC class II surface expression by regulated ubiquitination of a lysine residue in the β-chain. Little is known concerning how these enzymes target their specific substrates. Here we show that recognition of HLA-DR by MARCH proteins is complex. Several features associated with the transmembrane domain and bordering regions influence the overall efficiency of receptor internalization. A cluster of residues at the interface of the lipid bilayer and the cytosol plays the most important role in MARCH8 recognition of HLA-DRβ. Variation in this sequence also determines specificity of MARCH9 for HLA-DQ. Residues located in helical face four of HLA-DRβ together with a charged residue at the boundary with the stalk region also contribute significantly to recognition. Truncation analysis suggested that a dileucine-like motif in the DRβ cytoplasmic tail influences the efficiency of co-localization of HLA-DR with MARCH8. The DRβ-encoded acceptor lysine functioned optimally when placed in its natural location relative to the bilayer. In the DRα/DRβ dimer most other amino acids in the cytoplasmic tail could be substituted for alanine with minimal influence on function. Our data support a model whereby multiple features of HLA-DR are involved in substrate recognition by MARCH8. The single most important region is located at the interface between the transmembrane domain and the cytosol. Variation in sequence in this location between different class II isotypes controls efficiency of recognition by different MARCH E3 ligases.
authors: Division of Immunology, Department of Pathology, University of Cambridge, Cambridge CB2 1QP, United Kingdom.
authors: Martin
authors: M
authors: Jahnke
authors: John
authors: J
authors: Trowsdale
authors: Adrian P
authors: AP
authors: Kelly
mesh_terms: D020816:Amino Acid Motifs; D000818:Animals; D005786:Gene Expression Regulation / Q000502:physiology*; D057809:HEK293 Cells; D006683:HLA-DQ Antigens / Q000235:genetics / Q000378:metabolism*; D059805:HLA-DR alpha-Chains / Q000235:genetics / Q000378:metabolism*; D059806:HLA-DR beta-Chains / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D008565:Membrane Proteins / Q000235:genetics / Q000378:metabolism*; D051379:Mice; D017434:Protein Structure, Tertiary; D044767:Ubiquitin-Protein Ligases / Q000235:genetics / Q000378:metabolism*
chemical_list: D006683:HLA-DQ Antigens; D059805:HLA-DR alpha-Chains; D059806:HLA-DR beta-Chains; D008565:Membrane Proteins; C568898:MARCHF8 protein, human; C517365:MARCHF9 protein, human; D044767:Ubiquitin-Protein Ligases