Identification and analysis of the human CD160 promoter: implication of a potential AML-1 binding site in promoter activation.
CD160 is a glycosylphosphatidylinositol-anchored multimer expressed at the cell surface of subsets of cytotoxic T-lymphocytes and natural killer cells. Although CD160 is an important molecule for the control of cell-mediated cytotoxic responses, the mechanisms of regulation of its expression is unknown. We investigated the regulation of CD160 transcription by localizing and analyzing its promoter. The CD160 gene is encoded on chromosome 1, contains 6 exons with the translation initiation codon in exon 3. Bioinformatics analysis pointed to three potential promoter regions, two upstream of exon 1 and one in front of exon 3. RACE-PCR analysis identified a single transcription start site (TSS) and reporter gene transfections localized the active region immediately upstream of exon 1. Sequential deletion analysis led to the identification of a 371 bp sequence, located between -314 and +57 relative to the TSS, as the core promoter sequence driving CD160 gene transcription. Sequence alignment of the mouse and human CD160 genomic promoter region revealed a strong homology in the 371 bp sequence identified and pointed out to three conserved transcription factor binding sites for acute myelogenous leukemia-1 (AML-1), FREAC-4 and Sox17. Site-directed mutagenesis showed that the predicted AML-1 site is essential for the regulation of CD160 gene expression.
Schmitt
C
C
INSERM, U976, Paris, France. christian.schmitt@inserm.fr
Ghazi
B
B
Bellier
F
F
Bensussan
A
A
D000818:Animals; D015703:Antigens, CD / Q000235:genetics*; D001483:Base Sequence; D001665:Binding Sites; D002878:Chromosomes, Human, Pair 1 / Q000235:genetics; D050676:Core Binding Factor Alpha 2 Subunit / Q000235:genetics / Q000378:metabolism*; D005091:Exons; D051858:Forkhead Transcription Factors / Q000235:genetics* / Q000378:metabolism; D058851:GPI-Linked Proteins; D006801:Humans; D019169:Jurkat Cells; D007694:Killer Cells, Natural / Q000378:metabolism; D051379:Mice; D008969:Molecular Sequence Data; D011401:Promoter Regions, Genetic; D011971:Receptors, Immunologic / Q000235:genetics*; D055760:SOXF Transcription Factors / Q000235:genetics* / Q000378:metabolism; D016415:Sequence Alignment; D015533:Transcriptional Activation
D015703:Antigens, CD; C114505:CD160 protein, human; D050676:Core Binding Factor Alpha 2 Subunit; C500800:FOXD1 protein, human; D051858:Forkhead Transcription Factors; D058851:GPI-Linked Proteins; C493728:RUNX1 protein, human; D011971:Receptors, Immunologic; C109188:SOX17 protein, human; D055760:SOXF Transcription Factors