The HLA-G antigen is specifically expressed on trophoblasts at the maternal-fetal interface, while expression of classical class I HLA-A, -B, -C products is repressed in this tissue. The transcriptional level of the HLA-G gene is high in trophoblast cells and in JEG-3 choriocarcinoma cells, is markedly reduced in blood cells, and is shown here to be undetectable in the YT2C2 NK cell line. In an attempt to understand molecular mechanisms controlling cell-specific transcriptional regulation of the HLA-G gene in these cells, we focused our study on protein interaction with a 244-bp region located over 1.1 kb from exon 1, which has been shown to direct HLA-G expression in transgenic mouse trophoblast. Three specific complexes were detected, two of which are found exclusively in cells showing HLA-G transcriptional activity. The YT2C2 nuclear extracts contain restricted DNA-binding activity of an additional factor which could correlate with repression of HLA-G transcription in these cells.