Mapping the indirect p53 gene regulatory network

绘制间接 p53 基因调控网络

• 批准号: 498394458
• 负责人: Privatdozent Dr. Martin Fischer
• 金额: --
• 依托单位: Leibniz-Institut für Alternsforschung - Fritz-Lipmann-Institut e.V. (FLI)
• 依托单位国家: 德国
• 项目类别: Research Grants
• 资助国家: 德国
• 项目状态: 未结题
• 来源: https://gepris.dfg.de/gepris/projekt/498394458?language=en
• 关键词: Mapping; indirect; p53; gene; regulatory

Cancer is a group of age-related diseases that leads to more than 1.2 million deaths per year in the EU, and the transcription factor p53 serves as a central suppressor of carcinogenesis. It controls cell proliferation and apoptosis by regulating a plethora of target genes. Despite intensive efforts in the past decade, it is still unclear how p53 regulates many of its target genes, what co-factors are necessary for up or down-regulation, and what their regulation contributes to the suppression of carcinogenesis. Our incomplete picture of the molecular basis of gene regulation by p53 remains a critical barrier to our overall understanding of tumor suppression. We hypothesize that p53 employs several factors mediating gene regulation and control cell viability to suppress cancer growth; and that these pathways in the p53 gene regulatory network contribute independent, distinct, and essential gene regulatory activities. Using a meta-analysis approach that we developed earlier, we identified the novel p53-RFX7 signaling pathway. We found that the transcription factor RFX7 governs a tumor suppressor gene network in response to p53 and stress. In the case of RFX7, publicly available data on its sibling RFX5 enabled us to identify RFX7 as a critical transcriptional regulator that functions downstream of p53. However, this rather lucky circumstance cannot conceal that the discovery of new transcriptional regulators in p53’s network is difficult when working with public data alone. To overcome the limitation of our previous approach to identify novel candidate factors in the p53 gene regulatory network, we established methods to identify uncharted transcription factors and generated data that enables an informed screening approach. We will use CAGE-seq data to pinpoint promoter regions that mediate p53-dependent gene regulation and combine it with an established workflow of DNA-affinity purification followed by mass spectrometry to identify transcription factors that specifically bind the regulatory DNA fragments. Candidate transcription factors identified from the informed, comprehensive screening approach will subsequently be validated and characterized. The functional characterization of novel transcriptional regulators that operate downstream of p53 will provide insights into their contribution to p53-regulated processes as well as to their effect on target genes and biological processes irrespective of p53. Together, the goal of this project is to identify uncharted transcription factors contributing to p53-dependent transcriptional regulation. Thus, the project contributes to a better understanding of the p53 gene regulatory network and helps to determine the roles of the identified transcription factors in tumor suppression. In addition to acquiring new candidates for therapeutic intervention in the specific case of p53 malfunction, the project also serves as a blueprint for the genome-wide elucidation of other transcriptional networks.

癌症是一组与年龄相关的疾病，在欧盟每年导致超过120万人死亡，而转录因子p53是癌症发生的中心抑制因子。它通过调节大量的靶基因来控制细胞增殖和凋亡。尽管在过去的十年中进行了大量的努力，但仍不清楚p53如何调节其许多靶基因，上调或下调所需的辅助因子以及它们的调节对抑制致癌作用的贡献。我们对p53基因调控的分子基础的不完整了解仍然是我们全面理解肿瘤抑制的关键障碍。我们假设p53采用几种因子介导基因调控和控制细胞活力，以抑制癌症的生长，并在p53基因调控网络中的这些途径有助于独立的，独特的，和必要的基因调控活动。使用我们早期开发的荟萃分析方法，我们确定了新的p53-RFX 7信号通路。我们发现，转录因子RFX 7管理的肿瘤抑制基因网络的p53和压力。在RFX 7的情况下，关于其兄弟RFX 5的公开数据使我们能够将RFX 7鉴定为在p53下游发挥作用的关键转录调节因子。然而，这种相当幸运的情况并不能掩盖在p53网络中发现新的转录调节因子是困难的，当单独使用公共数据时。为了克服我们以前在p53基因调控网络中识别新候选因子的方法的局限性，我们建立了识别未知转录因子的方法，并生成了能够实现知情筛选方法的数据。我们将使用CAGE-seq数据来确定介导p53依赖性基因调控的启动子区域，并将其与DNA亲和纯化的既定工作流程相结合，然后进行质谱分析，以识别特异性结合调控DNA片段的转录因子。从知情的，全面的筛选方法确定的候选转录因子将随后进行验证和表征。新的转录调控，p53下游的功能特性将提供洞察他们的贡献p53调节过程，以及他们对靶基因和生物过程的影响，而不论p53。总之，这个项目的目标是确定未知的转录因子有助于p53依赖的转录调控。因此，该项目有助于更好地理解p53基因调控网络，并有助于确定已鉴定的转录因子在肿瘤抑制中的作用。除了在p53功能障碍的特定情况下获得新的治疗干预候选人外，该项目还可以作为其他转录网络的全基因组阐明的蓝图。