OVINE INTERFERON-TAU REGULATES UTERINE HORMONE RECEPTOR

羊干扰素-TAU 调节子宫激素受体

• 批准号: 6520950
• 负责人: FULLER W BAZER
• 金额: $ 26.19万
• 依托单位: TEXAS A&M UNIVERSITY HEALTH SCIENCE CTR
• 依托单位国家: 美国
• 财政年份: 1996
• 资助国家: 美国
• 起止时间: 1996-03-01 至 2006-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6520950
• 关键词: biological signal transduction; endometrium; estrogen inhibitor; estrogen receptors; gel mobility shift assay; gene expression; genetic enhancer element; genetic library; genetic promoter element; genetic transcription; hormone regulation /control mechanism; interferons; molecular dynamics; nucleic acid sequence; oxytocin; polymerase chain reaction; pregnancy; receptor expression; recombinant proteins; reproductive system pharmacology; tissue /cell culture; transcription factor; transfection; trophoblast; uterus

DESCRIPTION: (Adapted from the applicant's abstract) This is a revised renewal application in which research is proposed to investigate further the cellular and molecular mechanisms whereby ovine interferon-tau (IFN-tau) regulates estrogen receptor (ERalpha) and oxytocin receptor (OTR) gene expression in the endometrial epithelium. IFN-tau is produced by trophectoderm of ruminant concept uses and signals pregnancy recognition by suppressing ERalpha and OTR gene expression in the endometrial epithelium to prevent oxytocin-induced luteolytic pulses of prostaglandin F2alpha. Coincident with suppression of these genes, IFN-tau also induces or up regulates transcription of several Type I IFN-regulated genes. These disparate effects of IFN-tau are mediated by triggering a transcription factor network involving activation of the signal transducers of transcription (STAT) family, primarily STATS 1alpha/beta and 2, and induction of IFN regulatory factors (IRF) that bind to specific response elements and either activate or repress transcription of target genes. Results from the current grant period indicate that IFN-tau acts on the endometrial epithelium in vivo to suppress transcription of both the ERalpha and OTR genes and up regulate IRF-1 and IRF-2 repressor. Cloning and analysis of the 5' flanking promoter region (2.8 kb) of the ovine ERalpha gene revealed five predicted IRF elements (IRF-E) that were functional in binding IRF-1 and IRF-2. The working hypothesis is that IFN-tau induces an IRF repressor, such as IRF-2, which binds to functional IRF-Es in the ERalpha promoter to inhibit transcription. It has been demonstrated that IFN-tau inhibits activity of the 2.8 kb oERalpha promoter in a transient transfection assay using an immortalized ovine lumenal epithelial (LE) cell line. A transfection analysis of the oERalpha promoter indicates that both IRF and STAT binding elements are important for inhibitory effects of IFN-tau. The proposed research will continue to delineate cellular and molecular mechanisms underlying the action of IFN-tau in regulating ERalpha and OTR gene expression in the ovine uterus. Specific aims are to: 1) clone, sequence and functionally analyze the promoter of the ovine OTR gene, 2) determine which IRF family members are induced by IFN-tau in the endometrial epithelium, 3) analyze effects of IFN-tau regulated IRFs on activity of the ovine ERalpha and OTR promoters, and 4) test the hypothesis that STATs activated by IFN-tau mediate, in part, the inhibitory effects of IFN-tau on ovine ERalpha and OTR promoter activity.

描述：（改编自申请人的摘要）这是修订后的续展 提出研究以进一步研究细胞的应用 绵羊干扰素 tau (IFN-tau) 调节的分子机制 雌激素受体（ERα）和催产素受体（OTR）基因表达 子宫内膜上皮。 IFN-tau 由反刍动物滋养外胚层产生 概念通过抑制 ERalpha 和 OTR 来使用和发出怀孕识别信号 子宫内膜上皮中的基因表达可预防催产素诱导 前列腺素 F2α 的黄体溶解脉冲。与镇压同时发生 这些基因，IFN-tau 还诱导或上调几种类型的转录 I IFN 调节基因。 IFN-tau 的这些不同作用是由以下因素介导的： 触发涉及信号激活的转录因子网络 转录转导子 (STAT) 家族，主要是 STATS 1α/β 和 2， 以及诱导与特定反应结合的干扰素调节因子（IRF） 元件并激活或抑制靶基因的转录。结果 从目前的资助期限来看，表明 IFN-tau 作用于子宫内膜 体内上皮抑制 ERalpha 和 OTR 基因的转录 并上调 IRF-1 和 IRF-2 阻遏蛋白。 5'的克隆和分析 绵羊 ERalpha 基因的侧翼启动子区域 (2.8 kb) 揭示了五个 预测在结合 IRF-1 和 IRF-2 方面发挥功能的 IRF 元件 (IRF-E)。 工作假设是 IFN-tau 诱导 IRF 阻遏物，例如 IRF-2， 它与 ERalpha 启动子中的功能性 IRF-E 结合以抑制 转录。已证明 IFN-tau 可抑制 使用瞬时转染测定中的 2.8 kb oERalpha 启动子 永生化绵羊腔上皮（LE）细胞系。转染分析 oERalpha 启动子的序列表明 IRF 和 STAT 结合元件都是 对 IFN-tau 的抑制作用很重要。拟议的研究将 继续描绘作用背后的细胞和分子机制 IFN-tau 在调节绵羊子宫中 ERα 和 OTR 基因表达中的作用。 具体目标是：1) 启动子的克隆、测序和功能分析 绵羊 OTR 基因，2) 确定哪些 IRF 家族成员是由 子宫内膜上皮中的IFN-tau，3）分析IFN-tau调节的作用 IRF 对绵羊 ERalpha 和 OTR 启动子活性的影响，以及 4) 测试 假设 STATs 被 IFN-tau 激活，部分介导抑制作用 IFN-tau 对绵羊 ERα 和 OTR 启动子活性的影响。