Investigation of Gene Regulation by NF-kappaB Transcription factors

描述（由申请人提供）：NF- kappab (NF-？B)二聚体转录因子家族调节多种生理过程，包括免疫和炎症反应、细胞增殖和存活。组合的NF-？B二聚体由五个家族成员组成；p50, p52, RelA (p65)， cRel和RelB。这些二聚体通过结合特定的目标基因DNA序列来调节转录。B站点。我们实验室的长期目标是了解NF-？B二聚体在基因调控中起特殊作用。为了实现这一目标，我们已经采用了从生物物理学到遗传学的各种方法。最近的研究强调了NF-?翻译后修饰的作用。B二聚体和二聚体特异性共激活因子在调节特定靶基因中的作用。本提案的重点是通过磷酸化和共激活子识别事件来检查转录调节。我们采用遗传学、基因组学、生物化学和生物物理学的方法来研究两种NF-？在人类疾病中发挥最重要作用的B家族成员：RelA和p52。在目的1中，实验旨在研究RelA:CBP/p300复合物的转录控制机制。一些报道已经证明了RelA磷酸化和CBP/p300结合在靶基因调控中的重要性。其他研究表明，这种改变/相互作用的影响不是全球性的。每种修饰/相互作用仅以刺激和细胞类型特异性的方式影响一部分基因的表达。在本研究中，我们将探讨RelA磷酸化的作用和CBP/p300的相互作用机制及其对基因调控的影响。在目的2中，生物物理和体内实验旨在研究癌症相关的p52:Bcl3复合物的转录控制。作为同二聚体，NF-？据报道，bp52和p50与Bcl3结合激活转录，因此它们在免疫器官发生和耐受中起关键作用。本目的目的是研究p52同型二聚体如何与Bcl3结合的分子基础及其相互作用的特异性，以及这些复合物是否靶向？B序列。公共卫生相关性：NF-？二聚体转录激活因子B家族调控大量基因，对维持正常细胞生理至关重要。本提案的重点是了解启动子和转录辅激活子识别的调控NF-？二聚体。

DESCRIPTION (provided by applicant): The NF-kappaB (NF-?B) family of dimeric transcription factors regulates diverse physiological processes including immune and inflammatory responses, cell proliferation and survival. The combinatorial NF-?B dimers are formed from five family members; p50, p52, RelA (p65), cRel and RelB. These dimers regulate transcription by binding to specific DNA sequences of target genes known as ?B sites. The long term goals of our laboratories are to understand how NF-?B dimers function specifically in gene regulation. To achieve that goal, we have pursued a variety of approaches ranging from the biophysical to the genetic. Recent studies have emphasized the role of post-translational modifications of NF-?B dimers and the role of dimer-specific co-activators in regulating specific target genes. The focus of this proposal is to examine transcriptional regulation by phosphorylation and coactivator recognition events. We employ genetic, genomic, biochemical and biophysical approaches to investigate the two NF-?B family members that play the most prominent roles in human disease: RelA and p52. In aim 1 experiments are designed to investigate the mechanisms of transcriptional control by the RelA:CBP/p300 complex. Several reports have demonstrated the importance of RelA phosphorylation, and CBP/p300 binding in target gene regulation. Other studies have revealed that the effect of this modification/interaction is not global. Each modification/interaction affects the expression of only a subset of genes in stimulus and cell type specific manner. In this study we will investigate the role of RelA phosphorylation and CBP/p300 interaction mechanism and its impact on gene regulation. In aim 2 Biophysical and in vivo experiments are designed to study the transcriptional control by the cancer-associated p52:Bcl3 complex. As homodimers, NF-?B p52 and p50 have been reported to associate with Bcl3 to activate transcription, and as such they play critical roles in immune organogenesis and tolerance. The goal of this aim is to investigate the molecular basis of how p52 homodimer associates with Bcl3 and the specificity of the interaction, and if these complexes target a subset of ?B sequences. Public Health Relevance: The NF-?B family of dimeric transcription activators, which regulates a large number of genes, is critical for maintaining normal cell physiology. The focus of this proposal is to understand the regulation of promoter and transcriptional coactivator recognition by the NF-?B dimers.