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Papers In Press, published online ahead of print March 23, 2000
Pathology, Yale University School of Medicine, New Haven, CT 06510
Corresponding Author: david.johnson{at}yale.edu
HLA class I molecule expression was investigated by DNA mediated gene transfer. Cell surface expression was increased up to 75% by transfection of HLA-A2 or HLA-B8 heavy chain genes but not genes encoding light chains (
J. Biol. Chem, 10.1074/jbc.M910035199
Submitted on December 18, 1999
Revised on March 22, 2000
Accepted on March 23, 2000
Dependence of Elevated HLA Class I Molecule Expression on Increased Heavy Chain, Light Chain (
2m), TAP, Tapasin, and Peptide
2m), TAP, or tapasin. IFN treatment further increased expression of transfected heavy chains, suggesting that IFN inducible molecules support heavy chain expression. IFN induces 2m, TAP and tapasin mRNAs. Transfected heavy chain expression increased upon cotransfection with genes encoding TAP1 and TAP2 but not individual TAP subunits, 2m or tapasin. Tetracycline-inducible heavy chain gene expression was also increased by IFN treatment or TAP cotransfection, suggesting that IFN-induced TAP supports heavy chain maturation. Expression of a mutant that does not interact strongly with TAP, HLA-A2-T134K, was also increased by IFN. Inhibition of TAP-dependent peptide transport by ICP47 reduced heavy chain expression. Expression of HLA-A2, but not HLA-B8, was restored in ICP47 cells by HLA-A2-binding (IP-30) signal peptides. However, these peptides did not further increase transfected HLA-A2 expression, suggesting that peptide availability does not limit heavy chain expression in the absence of ICP47. These results suggest that cytokine induced TAP supports maturation of HLA class I molecules through combined chaperone and peptide supply functions.
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