IDENTIFICATION OF FAB-7 BOUNDARY PROTEINS

FAB-7 边界蛋白的鉴定

• 批准号: 7957799
• 负责人: Paul D Schedl
• 金额: $ 0.33万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-09-01 至 2010-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7957799
• 关键词: Adult; Binding; Biology; Chromatin; Cis-Acting Sequence; Complex; Computer Retrieval of Information on Scientific Projects Database; DNA Sequence; Development; Distal; Drosophila genus; Elements; Embryo; Embryonic Development; Ensure; Eukaryota; Funding; Fungal Genome; Grant; Institution; Proteins; Research; Research Personnel; Resources; Side; Site; Source; Staging; Tissues; Trans-Activators; United States National Institutes of Health; fly; nuclease; protein complex

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. The Fab-7 boundary functions to ensure the autonomous activity of the iab-6 and iab-7 cis-regulatory domains in the Drosophila Bithorax Complex from early embryogenesis through to the adult stage. Although Fab-7 is only required for the proper development of a single posterior parasegment, it is active in all tissues and stages of development that have been examined. In this respect Fab-7 resembles conventional constitutive boundaries in flies and other eukaryotes that act through ubiquitous cis-elements and trans-acting factors. The Fab-7 boundary spans a DNA sequence of ~1.2 kb and contains 3 chromatin specific nuclease hypersensitive sites. We have examined the functioning of cis-acting sequences in the largest of these hypersensitive sites, HS1, in detail. Though Fab-7 boundary activity is constitutive, we found that HS1 is divided into sub-elements whose boundary activity is restricted to only a specific stage in development. One sub-element, which is located on the proximal side of HS1, is active only in the early embryo. A second sub-element, which is located in the middle of HS1, is not active in early embryos; however, it becomes active mid-way through embryogenesis and then remains active through to the adult stage. A third sub-element on the distal side of HS1 confers boundary activity from early through mid-embryogenesis but not later. Through this project, we identified candidate proteins that are likely to confer this early-stage specific boundary activity, and then found that these proteins make a heterogeneous protein complex to bind the target sequence. We also would like to identify the trans-acting factors that confer other stage specific boundary activity in each.

该子项目是利用该技术的众多研究子项目之一 资源由 NIH/NCRR 资助的中心拨款提供。子项目及 研究者 (PI) 可能已从 NIH 的另一个来源获得主要资金， 因此可以在其他 CRISP 条目中表示。列出的机构是 对于中心来说，它不一定是研究者的机构。 Fab-7 边界的功能是确保果蝇 Bithorax 复合体中 IAB-6 和 IAB-7 顺式调控结构域从早期胚胎发生到成年阶段的自主活动。 尽管 Fab-7 仅是单个后副节段正常发育所必需的，但它在已检查的所有组织和发育阶段中都很活跃。 在这方面，Fab-7 类似于果蝇和其他真核生物中通过普遍存在的顺式元件和反式作用因子发挥作用的传统组成边界。 Fab-7 边界跨越约 1.2 kb 的 DNA 序列，并包含 3 个染色质特异性核酸酶超敏位点。 我们详细研究了最大的过敏位点 HS1 中顺式作用序列的功能。 尽管 Fab-7 边界活性是组成型的，但我们发现 HS1 被分为多个子元素，其边界活性仅限于发育的特定阶段。 位于 HS1 近侧的一个子元件仅在早期胚胎中活跃。 第二个子元件位于 HS1 中部，在早期胚胎中不活跃；然而，它在胚胎发生的中期变得活跃，然后一直活跃到成年阶段。 HS1 远端的第三个子元件赋予从胚胎发生早期到中期的边界活性，但不包括后期。 通过这个项目，我们鉴定了可能赋予这种早期特定边界活性的候选蛋白质，然后发现这些蛋白质形成异质蛋白质复合物来结合目标序列。 我们还想确定赋予每个阶段其他阶段特定边界活动的反式作用因素。