Rewiring cell cycle-regulated transcription in response to stress

重新连接细胞周期调节的转录以应对压力

• 批准号: 9273541
• 负责人: Jennifer A Benanti
• 金额: $ 36万
• 依托单位: UNIV OF MASSACHUSETTS MED SCH WORCESTER
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-05-15 至 2020-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9273541
• 关键词: Address; Alleles; Binding; Biological Assay; Biological Process; Calcineurin; Cancer Etiology; Cations; Cell Cycle; Cell Cycle Progression; Cell Cycle Regulation; Cell Cycle Stage; Cell Wall; Cell division; Cells; Cellular Stress; Cellular Structures; Chromatin; Consensus; Coupled; Cyclin-Dependent Kinases; Data; Data Set; Dependency; Disease; Eukaryota; Event; Gene Expression; Gene Expression Profile; Gene Targeting; Genetic; Genetic Transcription; Genome Stability; Genomic approach; Goals; Growth; Individual; Measures; Modeling; Modification; Mutagenesis; Mutation; Normal Cell; Output; PPP3CA gene; Phosphorylation; Phosphorylation Site; Post-Translational Protein Processing; Proteins; Proteomics; Regulation; Regulatory Pathway; Reporter; Role; S Phase; Saccharomyces cerevisiae; Saccharomycetales; Signal Transduction; Site; Stress; System; Testing; Time; Transcript; Work; Yeasts; alkalinity; base; calcineurin phosphatase; cancer cell; deep sequencing; experimental study; fitness; insight; mutant; network models; prevent; programs; protein function; public health relevance; response; therapy development; transcription factor

 DESCRIPTION (provided by applicant) Regulated progression through the cell division cycle is important to maintain the stability of the genome and to prevent uncontrolled cell division. The steps of the cell cycle are ordered by an underlying transcriptional program, which coordinates the expression of genes with the times in the cycle when their functions are needed. This tightly regulated pattern of gene expression is disrupted in nearly all cancer cells, underscoring its importance. High-throughput genomic approaches have identified TF networks that control cell cycle- regulated gene expression in diverse eukaryotes, the best understood being the yeast Saccharomyces cerevisiae. The prevailing model suggests that key TFs at each stage of the cell cycle activate expression of downstream TFs to drive the cell cycle forward. However, this model does not accurately predict many of the downstream effects that are observed when individual TFs are inactivated or deleted. A greater understanding of the coordination between TF proteins in the network is needed to fully understand this fundamental mechanism of cell-cycle control. Here we will address this outstanding issue by characterizing the response of each cell cycle TF (and their downstream effectors) to defined environmental or genetic perturbations. In the first aim, we will elucidate the dynamics of changes in the expression, regulation, and activity of each TF in the network in response to environmental conditions that we have found alter TF phosphorylation by cyclin- dependent kinase (Cdk1). In the second aim, we will dissect the mechanisms underlying how multisite phosphorylation of each TF in the network impacts their functions. In addition, we will use systematic, saturating mutagenesis in combination with bulk competition assays to elucidate how phosphorylation of multiple residues within an unstructured region of a model TF is read out into a change in TF function. By examining multiple regulatory parameters of this TF network in detail, our work will lead to an integrated, mechanistic view of this network, and have a significant impact on our understanding of cell-cycle control.

 描述（由申请人提供） 通过细胞分裂周期调节的进展对于维持细胞周期的稳定性是重要的。 基因组并防止不受控制的细胞分裂。细胞周期的步骤由一个潜在的转录程序来安排，该程序协调基因的表达与周期中需要其功能的时间。这种严格调控的基因表达模式在几乎所有癌细胞中都被破坏，这突出了它的重要性。高通量基因组方法已经鉴定了在多种真核生物中控制细胞周期调节的基因表达的TF网络，最好理解的是酵母酿酒酵母。流行的模型表明，在细胞周期的每个阶段的关键TF激活下游TF的表达，以推动细胞周期向前发展。然而，该模型不能准确预测当单个TF失活或缺失时观察到的许多下游效应。需要更好地理解网络中TF蛋白之间的协调，以充分理解细胞周期控制的基本机制。在这里，我们将解决这个突出的问题，通过表征每个细胞周期TF（及其下游效应子）的反应，以确定环境或遗传扰动。在第一个目标中，我们将阐明网络中每个TF的表达、调节和活性的动态变化，以响应我们发现通过细胞周期蛋白依赖性激酶（Cdk 1）改变TF磷酸化的环境条件。在第二个目标中，我们将剖析网络中每个TF的多位点磷酸化如何影响其功能的机制。此外，我们将使用系统的，饱和诱变结合批量竞争试验，以阐明如何在一个模型TF的非结构化区域内的多个残基的磷酸化读出到TF功能的变化。通过详细研究这个TF网络的多个调节参数，我们的工作将导致这个网络的综合，机械的观点，并对我们理解细胞周期控制产生重大影响。