RUI: Negative Transcriptional Regulation of the Inducible Interleukin-4 Gene

RUI：诱导型 IL-4 基因的负转录调控

• 批准号: 9724032
• 负责人: Deborah Weiss
• 金额: $ 27.57万
• 依托单位: Williams College
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 1997
• 资助国家: 美国
• 起止时间: 1997-08-15 至 2001-07-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=9724032&HistoricalAwards=false
• 关键词: RUI; Negative; Transcriptional; Regulation; Inducible

9724032 Weiss For genes which exhibit strict tissue specific expression or are expressed only upon induction, the contribution by negative regulatory elements is particularly important. The objective of this research is to define the nuclear factors which interact with the negative regulatory element NEG-1 of the Interleukin-4 (IL-4) gene. IL-4 is produced only by mast cells and certain T cell subsets, and then only upon induction. Messenger RNA levels for IL-4 increase rapidly after induction and then decrease once the inducing stimulus is removed. This observation coupled with other data indicates that IL-4 production is under transcriptional control. Previous studies have demonstrated that there are several DNA regulatory elements within the 5' flanking region of this gene and that specific nuclear factors interact with these sequences to regulate the inducible transcription of IL-4. The recently defined DNA element NEG-1, (-326 to -302 relative to the transcription initiation start site) contributes to the down regulation of this gene. Preliminary results indicate that two sub-sequences (A and B) within NEG-1 interact co-operatively and that NEG-1 A forms a specific complex with mast cell nuclear factor(s) found in both unstimulated and stimulated cells. This research identify the nuclear factor(s) in direct contact with the A region of NEG-1, and will use heparin-sepharose and DNA affinity chromatography to purify enough factor to obtain sequence information. A cDNA library will be constructed and screened with degenerate oligonucleotides corresponding to the sequence information obtained. Positive clones will be subcloned, sequenced, and expressed; their NEG-1 A binding ability will be confirmed by EMSA. %%% For genes which exhibit strict tissue specific expression or are expressed only upon induction, the contribution by negative regulatory elements is particularly important. Interleukin-4 (IL-4) is produced only by mast cells and certain T cell subsets, and then only upon induction. The objective of this research is to define the nuclear factors which interact with the negative regulatory element NEG-1 of the IL-4 gene. ***

9724032 Weiss 对于表现出严格的组织特异性表达或仅在诱导后表达的基因，负调控元件的贡献尤其重要。 本研究的目的是确定与白细胞介素 4 (IL-4) 基因的负调控元件 NEG-1 相互作用的核因子。 IL-4 仅由肥大细胞和某些 T 细胞亚群产生，并且仅在诱导时产生。 IL-4 的信使 RNA 水平在诱导后迅速增加，然后在诱导刺激消失后下降。 这一观察结果与其他数据相结合表明 IL-4 的产生受到转录控制。 先前的研究表明，该基因的 5' 侧翼区域内存在多个 DNA 调控元件，并且特定的核因子与这些序列相互作用以调控 IL-4 的诱导转录。 最近定义的DNA元件NEG-1（相对于转录起始位点的-326至-302）有助于该基因的下调。 初步结果表明，NEG-1 内的两个子序列（A 和 B）协同相互作用，并且 NEG-1 A 与未刺激和刺激细胞中发现的肥大细胞核因子形成特定复合物。 本研究鉴定了与NEG-1 A区直接接触的核因子，并将使用肝素琼脂糖和DNA亲和层析纯化足够的因子以获得序列信息。 将构建cDNA文库，并用与所获得的序列信息相对应的简并寡核苷酸进行筛选。 阳性克隆将被亚克隆、测序和表达；它们的NEG-1A结合能力将由EMSA确认。 %%% 对于表现出严格的组织特异性表达或仅在诱导时表达的基因，负调控元件的贡献尤其重要。 白细胞介素 4 (IL-4) 仅由肥大细胞和某些 T 细胞亚群产生，且仅在诱导后产生。 本研究的目的是确定与 IL-4 基因的负调控元件 NEG-1 相互作用的核因子。 ***