title: A novel domain on HLA-DRbeta chain regulates the chaperone role of the invariant chain.
abstract: The human lymphocyte antigen (HLA) class II region encodes highly polymorphic peptide receptors, which associate in the ER to the chaperone invariant chain (Ii). Ii facilitates assembly of class II subunits to functional peptide receptors. We searched for a conserved structure on HLA-DR polypeptides that mediates contact to a previously identified proline-rich class-II-binding sequence of Ii. Major histocompatibility complex (MHC) class II beta chain sequences exhibit two conserved tryptophan residues separated by 22 amino acids. Inspection of this motif in the X-ray structure of DR3 showed TrpTyr residues in the vicinity of the Ii-derived fragment CLIP. Five DRbeta mutants were produced. Mutation at Tyr123, Trp153 and Asp152 residues abolished interaction to the proline-rich sequence of Ii. All mutants formed heterodimers with DRalpha, were capable of binding an antigenic sequence and were expressed on the cell surface of transfected cells. In the presence of endogenous DRbeta chain however, the TyrAspTrp mutant was not cell-surface exposed and did not co-isolate with Ii or DRalpha. The competition of the mutant with the endogenous DRbeta for binding to DRalpha indicates that a structure on DRbeta chain regulates assembly of DR subunits. Hence, the chaperone function of Ii is mediated through a conserved region on the beta2 domain of class II.
authors.affiliation: Division of Immunobiology, Institute of Genetics, University of Bonn, Römerstr. 164, 53117 Bonn, Germany.
authors.forename: Jürgen
authors.initials: J
authors.lastname: Neumann
authors.forename: Norbert
authors.initials: N
authors.lastname: Koch
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000944:Antigens, Differentiation, B-Lymphocyte / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D019556:COS Cells; D045744:Cell Line, Tumor; D002522:Chlorocebus aethiops; D018360:Crystallography, X-Ray; D019281:Dimerization; D006684:HLA-DR Antigens / Q000737:chemistry / Q000235:genetics / Q000502:physiology*; D000949:Histocompatibility Antigens Class II / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D006801:Humans; D015151:Immunoblotting; D047468:Immunoprecipitation; D008856:Microscopy, Fluorescence; D008958:Models, Molecular; D018832:Molecular Chaperones / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D008969:Molecular Sequence Data; D016296:Mutagenesis / Q000235:genetics; D009154:Mutation / Q000235:genetics; D016133:Polymerase Chain Reaction / Q000379:methods; D011392:Proline / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D011485:Protein Binding; D017386:Sequence Homology, Amino Acid; D014364:Tryptophan / Q000737:chemistry / Q000235:genetics / Q000378:metabolism
chemical_list: D000944:Antigens, Differentiation, B-Lymphocyte; D006684:HLA-DR Antigens; D000949:Histocompatibility Antigens Class II; D018832:Molecular Chaperones; C045084:invariant chain; D014364:Tryptophan; D011392:Proline