Involvement of clathrin and AP-2 in the trafficking of MHC class II molecules to antigen-processing compartments

Abstract

Major histocompatibility complex class II (MHC-II) molecules are composed of two polymorphic chains, α and β, which assemble with an invariant chain, li, in the endoplasmic reticulum. The assembled MHC-II complexes are transported to the Golgi complex and then to late endosomes/lysosomes, where li is degraded and αβ dimers bind peptides derived from exogenous antigens. Targeting of MHC-II molecules to these compartments is mediated by two dileucine-based signals in the cytoplasmic domain of li. These signals bind in vitro to two adaptor protein (AP) complexes, AP-1 and AP-2, which are components of clathrin coats involved in vesicle formation and cargo sorting. The physiological roles of these proteins in MHC-II molecule trafficking, however, remain to be addressed. Here, we report the use of RNA interference to examine the involvement of clathrin and four AP complexes (AP-1, AP-2, AP-3, and AP-4) in MHC-II molecule trafficking in vivo. We found that depletion of clathrin or AP-2 caused > 10-fold increases in li expression on the cell surface and a concomitant decrease in li localization to endosomal/lysosomal vesicles. In addition, depletion of clathrin or AP-2 delayed the degradation of li and reduced the surface expression of peptide-loaded αβ dimers. In contrast, depletion of AP-1, AP-3, or AP-4 had little or no effect. These findings demonstrate that clathrin and AP-2 participate in MHC-II molecule trafficking in vivo. Because AP-2 is only associated with the plasma membrane, these results also indicate that a significant pool of MHC-II molecules traffic to the endosomal-lysosomal system by means of the cell surface.