Mechanisms of function of tapasin, a critical major histocompatibility complex class I assembly factor.
For their efficient assembly in the endoplasmic reticulum (ER), major histocompatibility complex (MHC) class I molecules require the specific assembly factors transporter associated with antigen processing (TAP) and tapasin, as well as generic ER folding factors, including the oxidoreductases ERp57 and protein disulfide isomerase (PDI), and the chaperone calreticulin. TAP transports peptides from the cytosol into the ER. Tapasin promotes the assembly of MHC class I molecules with peptides. The formation of disulfide-linked conjugates of tapasin with ERp57 is suggested to be crucial for tapasin function. Important functional roles are also suggested for the tapasin transmembrane and cytoplasmic domains, sites of tapasin interaction with TAP. We show that interactions of tapasin with both TAP and ERp57 are correlated with strong MHC class I recruitment and assembly enhancement. The presence of the transmembrane/cytosolic regions of tapasin is critical for efficient tapasin-MHC class I binding in interferon-gamma-treated cells, and contributes to an ERp57-independent mode of MHC class I assembly enhancement. A second ERp57-dependent mode of tapasin function correlates with enhanced MHC class I binding to tapasin and calreticulin. We also show that PDI binds to TAP in a tapasin-independent manner, but forms disulfide-linked conjugates with soluble tapasin. Thus, full-length tapasin is important for enhancing recruitment of MHC class I molecules and increasing specificity of tapasin-ERp57 conjugation. Furthermore, tapasin or the TAP/tapasin complex has an intrinsic ability to recruit MHC class I molecules and promote assembly, but also uses generic folding factors to enhance MHC class I recruitment and assembly.
Rizvi
Syed Monem
SM
Department of Microbiology and Immunology, University of Michigan Medical School, 5641 Medical Science Building II, Ann Arbor, MI 48109-5620, USA.
Raghavan
Malini
M
D018528:ATP-Binding Cassette Transporters / Q000378:metabolism*; D000818:Animals; D016466:CHO Cells; D037282:Calreticulin / Q000378:metabolism; D006224:Cricetinae; D003412:Cricetulus; D004721:Endoplasmic Reticulum / Q000378:metabolism; D006367:HeLa Cells; D015395:Histocompatibility Antigens Class I / Q000378:metabolism*; D006801:Humans; D007371:Interferon-gamma / Q000378:metabolism; D026901:Membrane Transport Proteins / Q000378:metabolism*; D011485:Protein Binding; D019704:Protein Disulfide-Isomerases / Q000378:metabolism; D017510:Protein Folding
D018528:ATP-Binding Cassette Transporters; D037282:Calreticulin; D015395:Histocompatibility Antigens Class I; D026901:Membrane Transport Proteins; C107762:tapasin; C477327:transporter associated with antigen processing (TAP); D007371:Interferon-gamma; D019704:Protein Disulfide-Isomerases