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J. Biol. Chem., Vol. 275, Issue 22, 16643-16649, June 2, 2000
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2-Microglobulin), Transporter Associated with
Antigen Processing, Tapasin, and Peptide*
and
From the Department of Pathology, Yale University School of
Medicine, New Haven, Connecticut 06510
Human leukocyte antigen (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
(
2-microglobulin (2m)), transporter
associated with antigen processing (TAP), or tapasin. Interferon (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.
To whom correspondence should be addressed: 454 Boyer Center for
Molecular Medicine, Yale Medical School, 295 Congress Ave., New Haven,
CT 06510. Tel.: 203-737-2298; Fax: 203-737-2293; E-mail: david.johnson@yale.edu.
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