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Generation of a functional, soluble tapasin protein from an alternatively spliced mRNA

Genes & Immunity volume 5, pages 101–108 (2004)Cite this article

Abstract

The loading of newly synthesised MHC class I molecules (MHCI) with peptides requires the involvement of several endoplasmic reticulum (ER)-resident cofactors including calnexin, calreticulin, transporter associated with antigen processing, ERp57 and tapasin. In the absence of tapasin, MHC I complexes are loaded with suboptimal peptides and their recognition by cytotoxic T cells raised to high-affinity, immunodominant peptide epitopes is impaired. Here, we describe the cloning and functional assessment of an alternative spliced form of tapasin. From the EST database, we obtained a partially spliced tapasin cDNA that retained introns 4–6. When transfected into the tapasin-deficient cell line 0.220, the cDNA produced an alternatively spliced tapasin transcript that contained intron 5 (74 bp). This introduced a new stop codon that terminated translation immediately before the putative transmembrane domain and led to a tapasin molecule containing the lumenal domain plus 8 extra novel amino acids at its C-terminus. This molecule promoted peptide loading of HLA-B5 in 0.220 cell line, and restored normal HLA-B5 surface expression. However, the peptides loaded onto HLA-B5 were suboptimal compared to those loaded onto HLA-B5 in the presence of wild-type tapasin.

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Acknowledgements

We would like to thank G Screaton for helpful discussion. This work was supported by the Wellcome Trust. APW is a Wellcome Clinical Training Fellow.

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Authors and Affiliations

  1. Rheumatology Unit, Institute of Child Health, University College, London, UK

    B Gao

  2. Cancer Sciences Division, University of Southampton School of Medicine, Southampton, UK

    A Williams & T Elliott

  3. Nuffield Department of Clinical Medicine, University of Oxford, John Radcliffe Hospital, UK

    A Sewell

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  1. B Gao
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  2. A Williams
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  3. A Sewell
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  4. T Elliott
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Correspondence to T Elliott.

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Gao, B., Williams, A., Sewell, A. et al. Generation of a functional, soluble tapasin protein from an alternatively spliced mRNA. Genes Immun 5, 101–108 (2004). https://doi.org/10.1038/sj.gene.6364043

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