Molecular cloning of cDNAs for transcriptional factors of indoleamine 2,3-dioxygenase induced by IFN-gamma

IFN-γ诱导的吲哚胺2,3-双加氧酶转录因子cDNA的分子克隆

• 批准号: 06680636
• 负责人: TAKIKAWA Osamu
• 金额: $ 1.47万
• 依托单位: Osaka Bioscience Institute
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (C)
• 财政年份: 1994
• 资助国家: 日本
• 起止时间: 1994 至 1995
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-06680636/
• 关键词: STAT 1; interferon-gamma; indoleamine 2,3-dioxygenase; transcriptional factor; IRF-1; tryptophan metabolism; cytokine; ISRE; 分子生物学

The purpose of this study is to characterize the transcriptional factors involved in the IFN-gamma-mediated induction of indoleamine 2,3-dioxygenase (IDO), a tryptophan degrading enzyme. Until 1994, we have demonstrated that two IDO mRNAs, 1.7kb and 2.3kb, whch differ in size of 5' franking region are induced by IFN-gamma in all human cultured cell lines examined. Kinetics of both mRNA induction and analyzes of the 5' control region of IDO gene expression have shown that 1) induction of both mRNAs are blocked by inhibitors for protein synthesis, indicating the need for a de novo synthesis of protein factor (s), 2) the protein synthesis occurs within 3 h after the addition of IFN-gamma, 3) the 5' region of 1.7kb mRNA that contains IRF-1 element, ISRE element, a Y-box, and a X-box but has no IFN-gamma-responsive promoting activity for transcription of IDO mRNA,and 4) the 5' region of 2.3kb mRNA has an ISRE and a X-box with the IFN-gamma responsive enhancing activity.In 1995, we have examined the presence of regulatory protein (s) for the enhance element for transcription of 2.3kb IDO mRNA in nuclear or cytoplasmic extract of human lung fibroblasts (HEL), which exhibited the highest induction of IDO by the addition of IFN-gamma. However, we have not yet detect such factor (s) in the nuclear extract. On the other hand, Gupta et al [J Interferon Res.15 : 517-526 (1995)] have reported that both STAT1 and IRF-1 are involved in the transcription of IDO mRNA.To confirm this, analysis of IDO induction by IFN-gamma in STAT1 and IRF-1 deficient mice is in progress.

本研究的目的是鉴定参与 IFN-γ 介导的吲哚胺 2,3-双加氧酶 (IDO)（一种色氨酸降解酶）诱导的转录因子。直到1994年，我们已经证明，在所有检测的人类培养细胞系中，IFN-γ可以诱导两种IDO mRNA，1.7kb和2.3kb，其5'标记区域的大小不同。 mRNA 诱导动力学和 IDO 基因表达 5' 控制区分析表明：1) 两种 mRNA 的诱导均被蛋白质合成抑制剂阻断，表明需要从头合成蛋白质因子，2) 蛋白质合成在添加 IFN-gamma 后 3 小时内发生，3) 包含 IRF-1 元件的 1.7kb mRNA 5' 区域， ISRE元件、Y-盒和X-盒，但对IDO mRNA的转录没有IFN-γ响应性促进活性，并且4)2.3kb mRNA的5'区具有具有IFN-γ响应性增强活性的ISRE和X-盒。1995年，我们检查了细胞核或细胞核中2.3kb IDO mRNA转录的增强元件的调节蛋白的存在。 人肺成纤维细胞 (HEL) 的细胞质提取物，通过添加 IFN-γ，表现出最高的 IDO 诱导能力。然而，我们尚未在核提取物中检测到此类因子。另一方面，Gupta等人[J Interferon Res.15：517-526(1995)]报道STAT1和IRF-1都参与IDO mRNA的转录。为了证实这一点，正在对STAT1和IRF-1缺陷小鼠中IFN-γ诱导IDO进行分析。

项目成果

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