An extra molecule in addition to human tapasin is required for surface expression of beta2m linked HLA-B4402 on murine cell.
Murine MHC class I can be readily expressed on the surface of human cell lines, but human class I molecules are expressed on mouse cells at a reduced level. Both human beta-2-microglobulin (beta(2)m) and tapasin (Tpn) have been demonstrated to be required for proper human MHC class I surface expression. Here we report that besides beta(2)m and tapasin, an extra unidentified component is also critical for the expression of certain human class I alleles. By covalently linking HLA-B4402 heavy chain to beta(2)m (beta(2)m-B44) a pre-assembled class I molecule has been created, which can be efficiently expressed and travel to the surface in human cells. In spite of being able to express inside cells, the linked beta(2)m-B44 molecule does not express on the surface of a murine fibroblast. Further investigation shows that lack of appearance on the surface is not due to quick degradation of unloaded class I, since provision of HLA-B4402 binding peptide could not rescue impaired surface expression. Co-expression with human tapasin does not rescue the defect excluding tapasin as the critical component for expression and indicating that a novel component of human origin is required for efficient surface expression of beta(2)m-B44 in murine cells. Surprisingly, not only did the beta(2)m-B44 construct fail to express on murine cells but also the surface expression of native murine MHC class I Kb was greatly reduced in transfected cells. It is likely that the expressed linked chain competitively associates with a component of class I processing in murine cells, reducing the exit rate of assembled mouse class I molecules. The results together suggest an unknown mechanism, which leads to the trapping of class I molecules in the ER.
Flutter
Barry
B
Rheumatology Unit, Infection and Immunity, Institute of Child Health, University College London, United Kingdom.
Fu
Hong-Mei
HM
Wedderburn
Lucy
L
Gao
Bin
B
D000818:Animals; D045744:Cell Line, Tumor; D019281:Dimerization; D005347:Fibroblasts / Q000187:drug effects / Q000276:immunology; D015235:HLA-B Antigens / Q000276:immunology / Q000378:metabolism*; D006801:Humans; D026901:Membrane Transport Proteins / Q000276:immunology / Q000378:metabolism*; D051379:Mice; D010455:Peptides / Q000494:pharmacology; D017510:Protein Folding; D021381:Protein Transport; D013696:Temperature; D001613:beta 2-Microglobulin / Q000276:immunology / Q000378:metabolism*
D015235:HLA-B Antigens; D026901:Membrane Transport Proteins; D010455:Peptides; D001613:beta 2-Microglobulin; C107762:tapasin