McCutcheon J A, Gumperz J, Smith K D, Lutz C T, Parham P
Department of Structural Biology, Stanford University School of Medicine, California 94305, USA.
J Exp Med. 1995 Jun 1;181(6):2085-95. doi: 10.1084/jem.181.6.2085.
In comparison with HLA-A and -B, the protein products of the HLA-C locus are poorly characterized, in part because of their low level of expression at the cell surface. Here, we examine how protein-protein interactions during assembly and regulation of the mRNA level affect cell surface expression of HLA-C. We find that intrinsic properties of the HLA-C heavy chain proteins do not correlate with low cell surface expression: HLA-C heavy chains associate and dissociate with beta 2-microglobulin (beta 2m) at rates comparable to those found for HLA-A and -B, and increased competition for beta 2m does not alter the surface expression of HLA-C. From studies of chimeric genes spliced from the HLA-B7 and -Cw3 genes, we find that chimeric proteins containing the B7 peptide-binding groove can have low cell surface expression, suggesting that inefficiency in binding peptides is not the cause of low cell surface expression for HLA-C. The surface levels of HLA-A, -B, or -C in cells transfected with cDNA can be similar, implicating noncoding regions of HLA-C heavy chain genes in the regulation of surface expression. We find that HLA-C mRNA is expressed at lower levels than HLA-B mRNA and that this difference results from faster degradation of the HLA-C message. Experiments examining chimeric B7/Cw3 and B7/Cw6 genes suggest that a region determining low expression of HLA-C is to be found between the 3' end of exon 3 and a site in the 3' untranslated region, approximately 600 bases downstream of the translation stop codon.
与HLA - A和 - B相比,HLA - C基因座的蛋白质产物的特征了解较少,部分原因是它们在细胞表面的表达水平较低。在此,我们研究了mRNA水平的组装和调控过程中的蛋白质 - 蛋白质相互作用如何影响HLA - C的细胞表面表达。我们发现HLA - C重链蛋白的内在特性与低细胞表面表达不相关:HLA - C重链与β2 - 微球蛋白(β2m)结合和解离的速率与HLA - A和 - B相当,并且对β2m竞争的增加不会改变HLA - C的表面表达。通过对从HLA - B7和 - Cw3基因拼接的嵌合基因的研究,我们发现含有B7肽结合槽的嵌合蛋白可能具有低细胞表面表达,这表明肽结合效率低下不是HLA - C低细胞表面表达的原因。用cDNA转染的细胞中HLA - A、 - B或 - C的表面水平可能相似,这表明HLA - C重链基因的非编码区参与表面表达的调控。我们发现HLA - C mRNA的表达水平低于HLA - B mRNA,并且这种差异是由于HLA - C信息更快的降解所致。检查嵌合B7 / Cw3和B7 / Cw6基因的实验表明,决定HLA - C低表达的区域位于外显子3的3'末端与3'非翻译区中翻译终止密码子下游约600个碱基处的一个位点之间。