Lack of tyrosine 320 impairs spontaneous endocytosis and enhances release of HLA-B27 molecules.
Several lines of evidence suggest that endocytosis of MHC class I molecules requires conserved motifs within the cytoplasmic domain. In this study, we show, in the C58 rat thymoma cell line transfected with HLA-B27 molecules, that replacement of the highly conserved tyrosine (Tyr320) in the cytoplasmic domain of HLA-B27 does not hamper cell surface expression of beta2-microglobulin H chain heterodimers or formation of misfolded molecules. However, Tyr320 replacement markedly impairs spontaneous endocytosis of HLA-B27. Although wild-type molecules are mostly internalized via endosomal compartments, Tyr320-mutated molecules remain at the plasma membrane in which partial colocalization with endogenous transferrin receptors can be observed, also impairing their endocytosis. Finally, we show that Tyr320 substitution enhances release of cleaved forms of HLA-B27 from the cell surface. These studies show for the first time that Tyr320 is most likely part of a cytoplasmic sorting motif involved in spontaneous endocytosis and shedding of MHC class I molecules.
Institute for Molecular and Cell Biology, Porto, Portugal.
Susana G
SG
Santos
Antony N
AN
Antoniou
Paula
P
Sampaio
Simon J
SJ
Powis
Fernando A
FA
Arosa
D020816:Amino Acid Motifs / Q000235:genetics / Q000276:immunology; D019943:Amino Acid Substitution / Q000235:genetics; D000818:Animals; D001692:Biological Transport / Q000235:genetics / Q000276:immunology; D045744:Cell Line, Tumor; D002462:Cell Membrane / Q000378:metabolism; D004705:Endocytosis / Q000235:genetics* / Q000276:immunology; D015796:HLA-B27 Antigen / Q000235:genetics* / Q000378:metabolism*; D006801:Humans; D016297:Mutagenesis, Site-Directed; D017354:Point Mutation; D051381:Rats; D011990:Receptors, Transferrin / Q000378:metabolism; D014443:Tyrosine / Q000172:deficiency* / Q000235:genetics*; D001613:beta 2-Microglobulin / Q000378:metabolism
D015796:HLA-B27 Antigen; D011990:Receptors, Transferrin; D001613:beta 2-Microglobulin; D014443:Tyrosine