A single bottleneck in HLA-C assembly

Leonardo Sibilio, Aline Martayan, Andrea Setini, Elisa Lo Monaco, Elisa Tremante, Richard H. Butler, Patrizio Giacomini

Research output: Contribution to journalArticle

Abstract

Poor assembly of class I major histocompatibility HLA-C heavy chains results in their intracellular accumulation in two forms: free of and associated with their light chain subunit (β2-microglobulin). Both intermediates are retained in the endoplasmic reticulum by promiscuous and HLA-dedicated chaperones and are poorly associated with peptide antigens. In this study, the eight serologically defined HLA-C alleles and the interlocus recombinant HLA-B46 allele (sharing the HLA-C-specific motif KYRV at residues 66-76 of the α1-domain α-helix) were compared with a large series of HLA-B and HLA-A alleles. Pulse-labeling experiments with HLA-C transfectants and HLA homozygous cell lines demonstrated that KYRV alleles accumulate as free heavy chains because of both poor assembly and post-assembly instability. Reactivity with antibodies to mapped linear epitopes, co-immunoprecipitation experiments, and molecular dynamics simulation studies additionally showed that the KYRV motif confers association to the HLA-dedicated chaperones TAP and tapasin as well as reduced plasticity and unfolding in the peptide-binding groove. Finally, in vitro assembly experiments in cell extracts of the T2 and 721.220 mutant cell lines demonstrated that HLA-Cw1 retains the ability to form a peptide-receptive interface despite a lack of TAP and functional tapasin, respectively. In the context of the available literature, these results indicate that a single locus-specific biosynthetic bottleneck renders HLA-C peptide-selective (rather than peptide-unreceptive) and a preferential natural killer cell ligand.

Original languageEnglish
Pages (from-to)1267-1274
Number of pages8
JournalJournal of Biological Chemistry
Volume283
Issue number3
DOIs
Publication statusPublished - Jan 18 2008

ASJC Scopus subject areas

  • Biochemistry

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    Sibilio, L., Martayan, A., Setini, A., Monaco, E. L., Tremante, E., Butler, R. H., & Giacomini, P. (2008). A single bottleneck in HLA-C assembly. Journal of Biological Chemistry, 283(3), 1267-1274. https://doi.org/10.1074/jbc.M708068200