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Localization of upstream DNA regions regulating T cell immunoglobulin and mucin domain-3 expression in CD4+ T cells

Yun, Su-jin; Jun, Ka-jung; Komori, Kuniharu; Park, Sun
Department of Microbiology, Ajou University School of Medicine
T cell immunoglobulin and mucin domain-3 (TIM-3) is expressed in various immune cells and regulates immune responses of these cells. In CD4+ T cells, TIM-3 expression is induced by activation signals especially in Th1 differentiation condition. In this study we localized regulatory regions in upstream DNA sequences of TIM-3 coding gene. We found the region of -144 bp ~ -1 bp from transcription start site of TIM-3 as a minimal promoter. This region contains a putative AP-1 binding site and its mutation reduced luciferase reporter activity. When AP-1 is overexpressed, both TIM-3 proximal promoter activity and mRNA expression were increased. Among upstream 4 different CNS, increased dimethylation of histone H3 lysin4 (H3K4me2) indicating enhancer region was observed on CNS4 in Jurkat T cells and Th1 cells expressing TIM-3 but not in Th2 cells not expressing TIM-3. Furthermore, CNS4 increased TIM-3 minimal promoter activity only when two putative NF-κB binding sites in this region was not mutated. Taken together, our results suggest that minimal TIM-3 promoter region containing an AP-1 binding site and CNS4 containing NF-κB binding sites are important in TIM-3 transcriptional regulation.
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