Negative transcriptional regulatory mechanisms of cytokine LD78 which regulates hematopoiesis

造血细胞因子LD78的负转录调控机制

• 批准号: 06670149
• 负责人: NOMIYAMA Hisayuki
• 金额: $ 1.15万
• 依托单位: KUMAMOTO UNIVERSITY
• 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-06670149/
• 关键词: Cytokine; Chemokine; LD78; Transcriptional factors; Silencer; MCP-3; YAC; Database

From several lines of evidence, it seems likely that a silencer element inhibits chemokine LD78 gene expression in the cells where no message is found. We have tried to identify and purify the transcriptional regulators which bind to this silencer element and to the ICK-1 element. The ICK-1 element is present in the promoter and regulates the LD78 transcription positively and negatively. However, These transcriptional regulators are very unstable and therefore it was difficult to purify them. We also analyzed the promoter of a chemokine family member MCP-3. MCP-3 promoter was active in HeLa cells, although no MCP-3 message was found. Since chomkines genes are clustered on human chromosome 17, the silencer element which inhibits the LD78 expression may also repress the expression of MCP-3 and other chemokine genes. We therefore constructed a yeast artificial chromosome (YAC) contig of several Mb long containing all chemokine genes present on chromosome 17q11.2. The contig also contains four chemokine-like genes which were deposited in GenBank as ESTs. This YAC contig is useful for identification of new chemokine genes in addition to the analysis of the silencer element.

从几条证据来看，似乎有可能是一种沉默元件抑制了趋化因子LD78基因在没有发现信息的细胞中的表达。我们试图鉴定和纯化与这个沉默元件和ick-1元件结合的转录调控因子。ICK-1元件存在于启动子中，对LD78的转录有正向和负向调节作用。然而，这些转录调控因子非常不稳定，因此很难纯化。我们还分析了趋化因子家族成员MCP-3的启动子。MCP-3启动子在HeLa细胞中有活性，但未发现MCP-3信息。由于Chomkines基因聚集在人类17号染色体上，抑制LD78表达的沉默元件也可能抑制MCP-3和其他趋化因子基因的表达。因此，我们构建了一个酵母人工染色体(YAC)重叠群，由几个Mb Long组成，包含染色体17q11.2上存在的所有趋化因子基因。该重叠群还包含四个趋化素样基因，这些基因作为EST保存在GenBank中。该YAC重叠群除了用于沉默元件的分析外，还可用于鉴定新的趋化因子基因。

项目成果

K.Naruse et al: "A YAC contig of the human CC chemokine genes clustered on chromosome 17q11.2" Genomics. (in press). (1996)

K.Naruse 等人：“人类 CC 趋化因子基因的 YAC 重叠群聚集在染色体 17q11.2 上”基因组学。

K.Naruse et al.: "A YAC contig of the human CC chemokine genes clustered on chromosome 17q11.2" Genomics. (in press). (1996)

K.Naruse 等人：“人类 CC 趋化因子基因的 YAC 重叠群聚集在染色体 17q11.2 上”基因组学。