Novel aspects of degradation of T cell receptor subunits from the endoplasmic reticulum (ER) in T cells: importance of oligosaccharide processing, ubiquitination, and proteasome-dependent removal from ER membranes.
Expression of the T cell antigen receptor (TCR) on the surface of thymocytes and mature T cells is dependent on the assembly of receptor subunits into TCRs in the endoplasmic reticulum (ER) and their successful traversal of the secretory pathway to the plasma membrane. TCR subunits that fail to exit the ER for the Golgi complex are degraded by nonlysosomal processes that have been referred to as "ER degradation". The molecular basis for the loss of the TCR CD3-delta and TCR-alpha subunits from the ER was investigated in lymphocytes. For CD3-delta, we describe a process leading to its degradation that includes trimming of mannose residues from asparagine-linked (N-linked) oligosaccharides, generation of ubiquitinated membrane-bound intermediates, and proteasome-dependent removal from the ER membrane. When either mannosidase activity or the catalytic activity of proteasomes was inhibited, loss of CD3-delta was markedly curtailed and CD3-delta remained membrane bound in a complex with CD3-epsilon. TCR-alpha was also found to be degraded in a proteasome-dependent manner with ubiquitinated intermediates. However, no evidence of a role for mannosidases was found for TCR-alpha, and significant retrograde movement through the ER membrane took place even when proteasome function was inhibited. These findings provide new insights into mechanisms employed to regulate levels of TCRs, and underscore that cells use multiple mechanisms to target proteins from the ER to the cytosol for degradation.
Yang
M
M
Laboratory of Immune Cell Biology, Division of Basic Sciences, National Cancer Institute, National Institutes of Health, Bethesda, Maryland 20892-1152, USA.
Omura
S
S
Bonifacino
J S
JS
Weissman
A M
AM
D000818:Animals; D017252:CD3 Complex / Q000378:metabolism; D003546:Cysteine Endopeptidases / Q000502:physiology*; D004721:Endoplasmic Reticulum / Q000378:metabolism*; D008361:Mannosidases / Q000378:metabolism; D051379:Mice; D008810:Mice, Inbred C57BL; D008811:Mice, Inbred DBA; D009097:Multienzyme Complexes / Q000502:physiology*; D009844:Oligosaccharides / Q000378:metabolism*; D046988:Proteasome Endopeptidase Complex; D016693:Receptors, Antigen, T-Cell, alpha-beta / Q000378:metabolism*; D016692:Receptors, Antigen, T-Cell, gamma-delta / Q000378:metabolism*; D014452:Ubiquitins / Q000378:metabolism*
D017252:CD3 Complex; D009097:Multienzyme Complexes; D009844:Oligosaccharides; D016693:Receptors, Antigen, T-Cell, alpha-beta; D016692:Receptors, Antigen, T-Cell, gamma-delta; D014452:Ubiquitins; D008361:Mannosidases; D003546:Cysteine Endopeptidases; D046988:Proteasome Endopeptidase Complex