Unique Expression of Major Histocompatibility Complex Class I Proteins in the Absence of Glucose Trimming and Calnexin Association

Abstract

Recent evidence indicates that efficient expression of major histocompatibility complex (MHC) complexes requires their interaction with the resident endoplasmic reticulum (ER) chaperone calnexin, which for certain proteins functions as a lectin specific for monoglucosylated glycans. In the current report, we studied the expression of MHC class I proteins in BW wild type thymoma cells (BW WT) and glucosidase II-deficient BW PHAR2.7 cells. Consistent with a requirement for glucose (Glc) trimming for interaction of class I proteins with calnexin, we found that nascent H-2Kk proteins associated with calnexin in untreated BW WT cells, but not in BW WT cells treated with the glucosidase inhibitor castanospermine (cas), or in untreated glucosidase II-deficient BW PHAR2.7 cells. Surprisingly, we found that H-2Kk expression occurred with similar efficiency in BW PHAR2.7 cells as in BW WT cells and that formation of nascent H-2Kk complexes was perturbed by cas treatment in BW WT cells but not in BW PHAR2.7 cells. Finally, it was noted that expression of the molecular chaperone Bip was markedly increased in BW PHAR2.7 cells relative to BW WT cells, which is suggested to play a role in regulating the expression of H-2Kk complexes in BW PHAR2.7 cells. The current study demonstrates that Glc trimming is required for efficient interaction of nascent H-2Kk proteins with calnexin; that expression of MHC class I proteins can, under certain conditions, proceed effectively in the absence of Glc trimming and calnexin association; and that Bip expression is markedly increased under conditions where diglucosylated glycans persist on nascent glycoproteins within the ER. These data are consistent with the hypothesis that alternative oligomerization pathways exist for class I proteins within the quality control system of the ER that have differential requirements for removal of Glc residues from nascent glycan chains.