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* Section of Retroviral Immunology, Center for Biologics Evaluation and Research, Food and Drug Administration, Bethesda, MD 20892;
Laboratory of Molecular Endocrinology, Medstar Research Institute, Washington Hospital Center, Washington, DC 20010; and
Department of Bioregulation, National Institute of Infectious Diseases, Tokyo, Japan
To clarify the molecular basis of human TLR9 (hTLR9) gene expression, the activity of the hTLR9 gene promoter was characterized using the human myeloma cell line RPMI 8226. Reporter gene analysis and EMSA demonstrated that hTLR9 gene transcription was regulated via four cis-acting elements, cAMP response element, 5'-PU box, 3'-PU box, and a C/EBP site, that interacted with the CREB1, Ets2, Elf1, Elk1, and C/EBP
transcription factors. Other members of the C/EBP family, such as C/EBP
, C/EBP
, and C/EBP
, were also important for TLR9 gene transcription. CpG DNA-mediated suppression of TLR9 gene transcription led to decreased binding of the trans-acting factors to their corresponding cis-acting elements. It appeared that suppression was mediated via c-Jun and NF-
B p65 and that cooperation among CREB1, Ets2, Elf1, Elk1, and C/EBP culminated in maximal transcription of the TLR9 gene. These findings will help to elucidate the mechanism of TLR9 gene regulation and to provide insight into the process by which TLR9 evolved in the mammalian immune system.
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