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Influence of the tapasin C terminus on the assembly of MHC class I allotypes

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Abstract

Several endoplasmic reticulum proteins, including tapasin, play an important role in major histocompatibility complex (MHC) class I assembly. In this study, we assessed the influence of the tapasin cytoplasmic tail on three mouse MHC class I allotypes (H2-Kb, -Kd, and -Ld) and demonstrated that the expression of truncated mouse tapasin in mouse cells resulted in very low Kb, Kd, and Ld surface expression. The surface expression of Kd also could not be rescued by human soluble tapasin, suggesting that the surface expression phenotype of the mouse MHC class I molecules in the presence of soluble tapasin was not due to mouse/human differences in tapasin. Notably, soluble mouse tapasin was able to partially rescue HLA-B8 surface expression on human 721.220 cells. Thus, the cytoplasmic tail of tapasin (either mouse or human) has a stronger impact on the surface expression of murine MHC class I molecules on mouse cells than on the expression of HLA-B8 on human cells. A K408W mutation in the mouse tapasin transmembrane/cytoplasmic domain disrupted Kd folding and release from tapasin, but not interaction with transporter associated with antigen processing (TAP), indicating that the mechanism whereby the tapasin transmembrane/cytoplasmic domain facilitates MHC class I assembly is not limited to TAP stabilization. Our findings indicate that the C terminus of mouse tapasin plays a vital role in enabling murine MHC class I molecules to be expressed at the surface of mouse cells.

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Acknowledgments

This work was supported by NIH Grant GM57428 (to J.C.S.), by an NIH/NCI Training Grant T32 CA009476 Fellowship (to L.S.), and by UNMC Graduate Studies Fellowships (to L.S. and A.T.). Core facilities at the University of Nebraska Medical Center receive support from the NIH/NCI Cancer Center Support Grant P30CA036727 (to the Eppley Cancer Center) and the Nebraska Research Initiative. The UNMC DNA Sequencing Core Facility receives partial support from the NIH/NCRR INBRE Program P20 RR016469 Grant. We thank Dr. Ping Wang for the gift of the tapasin cDNAs, Drs. T. Hansen, A. Grandea, L. Van Kaer, T. Spies, and P. Cresswell for gifts of cell lines, and Dr. T. Hansen for gifts of antibodies. We also gratefully acknowledge the assistance of Briana Ormsbee and the personnel at the University of Nebraska Medical Center Cell Analysis Facility, Monoclonal Antibody Facility, and DNA Sequencing Core Facility.

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  1. Eppley Institute for Research in Cancer and Allied Diseases and Department of Biochemistry and Molecular Biology, University of Nebraska Medical Center, 986805 Nebraska Medical Center, Omaha, NE, 68198-6805, USA

    Laura C. Simone, Xiaojian Wang, Amit Tuli, Mary M. McIlhaney & Joyce C. Solheim

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  1. Laura C. Simone
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  2. Xiaojian Wang
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  3. Amit Tuli
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Correspondence to Joyce C. Solheim.

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Simone, L.C., Wang, X., Tuli, A. et al. Influence of the tapasin C terminus on the assembly of MHC class I allotypes. Immunogenetics 61, 43–54 (2009). https://doi.org/10.1007/s00251-008-0335-x

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