title: Tapasin enhances peptide-induced expression of H2-M3 molecules, but is not required for the retention of open conformers.
abstract: H2-M3 is a class Ib MHC molecule that binds a highly restricted pool of peptides, resulting in its intracellular retention under normal conditions. However, addition of exogenous M3 ligands induces its escape from the endoplasmic reticulum (ER) and, ultimately, its expression at the cell surface. These features of M3 make it a powerful and novel model system to study the potentially interrelated functions of the ER-resident class I chaperone tapasin. The functions ascribed to tapasin include: 1) ER retention of peptide-empty class I molecules, 2) TAP stabilization resulting in increased peptide transport, 3) direct facilitation of peptide binding by class I, and 4) peptide editing. We report in this study that M3 is associated with the peptide-loading complex and that incubation of live cells with M3 ligands dramatically decreased this association. Furthermore, high levels of open conformers of M3 were efficiently retained intracellularly in tapasin-deficient cells, and addition of exogenous M3 ligands resulted in substantial surface induction that was enhanced by coexpression of either membrane-bound or soluble tapasin. Thus, in the case of M3, tapasin directly facilitates intracellular peptide binding, but is not required for intracellular retention of open conformers. As an alternative approach to define unique aspects of M3 biosynthesis, M3 was expressed in human cell lines that lack an M3 ortholog, but support expression of murine class Ia molecules. Unexpectedly, peptide-induced surface expression of M3 was observed in only one of two cell lines. These results demonstrate that M3 expression is dependent on a unique factor compared with class Ia molecules.
authors.affiliation: Department of Genetics, Washington University School of Medicine, St. Louis, MO 63110, USA.
authors.forename: L
authors.initials: L
authors.lastname: Lybarger
authors.forename: Y Y
authors.initials: YY
authors.lastname: Yu
authors.forename: T
authors.initials: T
authors.lastname: Chun
authors.forename: C R
authors.initials: CR
authors.lastname: Wang
authors.forename: A G
authors.initials: AG
authors.lastname: Grandea
authors.forename: L
authors.initials: L
authors.lastname: Van Kaer
authors.forename: T H
authors.initials: TH
authors.lastname: Hansen
mesh_terms: D000071181:ATP Binding Cassette Transporter, Subfamily B, Member 2; D000071450:ATP Binding Cassette Transporter, Subfamily B, Member 3; D018528:ATP-Binding Cassette Transporters / Q000378:metabolism; D000276:Adjuvants, Immunologic / Q000172:deficiency / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D000818:Animals; D017920:Antiporters / Q000172:deficiency / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D002461:Cell Line, Transformed; D000939:Epitopes / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D006183:H-2 Antigens / Q000378:metabolism; D006367:HeLa Cells; D064532:Histocompatibility Antigen H-2D; D015395:Histocompatibility Antigens Class I / Q000096:biosynthesis* / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D006801:Humans; D007136:Immunoglobulins / Q000172:deficiency / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D007739:L Cells; D026901:Membrane Transport Proteins; D051379:Mice; D016297:Mutagenesis, Site-Directed; D010455:Peptides / Q000378:metabolism / Q000494:pharmacology*; D011485:Protein Binding / Q000235:genetics / Q000276:immunology; D011487:Protein Conformation; D014162:Transfection
chemical_list: D000071181:ATP Binding Cassette Transporter, Subfamily B, Member 2; D000071450:ATP Binding Cassette Transporter, Subfamily B, Member 3; D018528:ATP-Binding Cassette Transporters; D000276:Adjuvants, Immunologic; D017920:Antiporters; D000939:Epitopes; D006183:H-2 Antigens; C078177:H2-M3 antigen; D064532:Histocompatibility Antigen H-2D; D015395:Histocompatibility Antigens Class I; D007136:Immunoglobulins; D026901:Membrane Transport Proteins; D010455:Peptides; C073119:TAP1 protein, human; C486171:Tap1 protein, mouse; C489516:Tap2 protein, mouse; C107762:tapasin; C071692:TAP2 protein, human