STAT1-DEPENDENT TRANSCRIPTION IN AIRWAY EPITHELIAL CELLS

描述 （摘自申请人摘要）申请人提出， 上皮细胞提供重要的生化信号， 免疫和炎症。 为了支持这种可能性，他们发现 上皮细胞表达一组功能性免疫反应基因 （ICAM-1、IRF-1、TAP 1和RANTES）在干扰素-γ的控制下， (IFN-γ）信号转导途径。 例如，上皮细胞 细胞间粘附分子-1（ICAM-1）的表达介导白细胞 气道中的运输（粘附、迁移和滞留） 通过作为白细胞B2-整联蛋白的受体而作用于上皮细胞。 在这 在此背景下，他们已经获得了ICAM-1介导上皮免疫的证据 可允许"适当"免疫应答的细胞相互作用（例如， 呼吸道病毒感染）或"不适当的"反应（例如，气道 哮喘中的炎症）。本申请基于以下观察： ICAM-1在气道上皮细胞上的表达依赖于特定的 IFN-γ驱动的信号转导途径，其反过来依赖于 活化的转录因子Stat1与ICAM-1基因的结合 启动子区 除了这种DNA/蛋白质相互作用之外，Statl还 似乎与转录辅激活因子p3OO相互作用， 转录因子Sp1完全激活基因转录。 的目的 本申请旨在确定Statl的结构特征， 与p3OO和Sp1的相互作用以及这些相互作用在 激活ICAM-1和相关免疫应答基因的转录。 这 这些信息然后可以用于改变ICAM-1基因激活和气道 通过阻断增强子途径和增强的炎症反应 阻遏物途径（例如，Statl显性负突变与腺病毒 EIA蛋白表达）。 因此，具体目标是：一。确定结构基础， Stat1/p300相互作用在ICAM-1基因激活中的功能作用。 他们 将使用他们的原代培养人气道上皮细胞模型来定义 Stat1与转录辅激活因子p300之间的相互作用。 Statl和p300的结构-功能研究将在2015年完成。 定义蛋白质/蛋白质相互作用和基因转录的实验 对于野生型和突变形式的Statl和p300。 二. 定义 ICAM-1基因中Statl/Sl相互作用结构基础和功能作用 activation. 他们将利用相同的细胞模型来表征 转录协同作用所需的结构域 Statl和Spl。Stat1和Sp1的结构-功能研究将在 使用蛋白质/蛋白质、蛋白质/DNA和基因转录来完成 用于野生型和突变形式的Statl和Spl的实验。(End的 摘要）

DESCRIPTION (Adapted from applicants' abstract) The applicants propose that airway epithelial cells provide critical biochemical signals for regulating immunity and inflammation. In support of this possibility, they have found that epithelial cells express a functional set of immune-response genes (ICAM-1, IRF-1, TAP1, and RANTES) under the control of an interferon-gamma (IFN-,gamma) signal transduction pathway. For example, epithelial cell expression of intercellular adhesion molecule-1 (ICAM- 1) mediates leukocyte trafficking (adhesion, transmigration, and retention) in the airway epithelium by serving as a receptor for leukocyte B2-integrins. In that context, they have obtained evidence that ICAM-1 mediates epithelial-immune cell interactions that may allow for "appropriate" immune responses (e.g., to a respiratory virus infection) or "inappropriate" responses (e.g., airway inflammation in asthma ). This application is based on the observation that ICAM-1 expression on airway epithelial cells depends on a specific IFN-gamma-driven signal transduction pathway that relies in turn on the binding of the activated transcription factor Statl to the ICAM-1 gene promoter region. In addition to this DNA/protein interaction, Statl also appears to interact with the transcriptional coactivator p3OO and the transcription factor Sp1 to fully activate gene transcription. The aim of this application is to determine the structural features of Statl that allow for interaction with p3OO and Sp1 and the role of these interactions in activating transcription for ICAM-1 and related immune-response genes. This information may then be used to modify ICAM-1 gene activation and the airway inflammatory response through blockade of enhancer-pathways and potentiation of repressor-pathways (e.g., Statl dominant-negative mutation and adenoviral EIA protein expression). Accordingly, The specific aims are to: I. Define the structural basis and functional role of Statl/p3OO interaction in ICAM-1 gene activation. They will use their primary culture human airway epithelial cell model to define the interaction between Statl and the transcriptional coactivator p3OO. Structure-function studies of Statl and p3OO will be accomplished in experiments that define protein/protein interactions and gene transcription for wild-type and mutated forms of Statl and p3OO. II. Define the structural basis and functional role of Statl/Spl-interaction in ICAM-1 gene activation. They will take advantage of the same cell model to characterize the structural domains required for the transcriptional synergy between Statl and Spl. Structure-function studies of Stat1 and Sp1 will be accomplished using protein/protein, protein/DNA, and gene transcription experiments for wild-type and mutated forms of Statl and Spl. (End of Abstract)