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

Peter J McCormick, José A Martina, Juan S Bonifacino

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98 Citations (Scopus)

Abstract

Major histocompatibility complex class II (MHC-II) molecules are composed of two polymorphic chains, alpha and beta, which assemble with an invariant chain, Ii, in the endoplasmic reticulum. The assembled MHC-II complexes are transported to the Golgi complex and then to late endosomes/lysosomes, where Ii is degraded and alphabeta 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 Ii. 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 Ii expression on the cell surface and a concomitant decrease in Ii localization to endosomal/lysosomal vesicles. In addition, depletion of clathrin or AP-2 delayed the degradation of Ii and reduced the surface expression of peptide-loaded alphabeta 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.
Original languageEnglish
Pages (from-to)7910-5
Number of pages6
JournalProceedings of the National Academy of Sciences
Volume102
Issue number22
DOIs
Publication statusPublished - 31 May 2005

Keywords

  • Adaptor Protein Complex 2
  • Antigen Presentation
  • Antigens, Differentiation, B-Lymphocyte
  • Cathepsin D
  • Clathrin
  • Flow Cytometry
  • HeLa Cells
  • Histocompatibility Antigens Class II
  • Humans
  • Immunoblotting
  • Immunoprecipitation
  • Lysosomes
  • Microscopy, Fluorescence
  • Protein Transport
  • RNA Interference
  • RNA, Small Interfering
  • Signal Transduction

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