Tissue-specific Transcriptional Regulation of Human Leukosialin (CD43) Gene Is Achieved by DNA Methylation

The expression of human leukosialin (CD43), a major sialoglycoprotein on the surface of hematopoietic cells, is regulated in cell lineage-specific as well as differentiation stage-specific manners. We have shown previously that transcription from the TATA-less promoter is mediated by the transcription factor Sp1, which binds to repeats of a GGGTGG motif in the 5'-flanking sequence. This regulatory region is ubiquitously functional in mammalian cells, providing a high transcriptional potential. No cis-acting element responsible for the specificity of this gene expression was revealed by extensive studies using transient as well as stable expression systems. Here, we demonstrate that DNA methylation plays a key role in leukosialin gene expression. Southern blot analysis of genomic DNAs from various human cell lines with methylation-sensitive and -insensitive restriction enzymes showed a tight correlation between gene activity and demethylation state of the 5'-region of the leukosialin gene. Consistent results were obtained from the same analysis of genomic DNAs from various human tissues. In addition, in vitro DNA methylation of the 5'-region drastically reduced transcriptional activity in a transient expression system. These results indicate that DNA methylation around the 5'-region of the leukosialin gene is required to shut off a high level of transcription. Thus, the tissue-specific expression of the leukosialin gene is constitutively achieved by alteration of DNA methylation.