The Glucocorticoid Receptor as Signal Integrator: Studying All Drug Resistance

项目总结/摘要 蛋白质位于信号通路的节点，它们的任务是整合这些信号以指导特定的 输出-一个是定制的细胞的需要在当时。Miles Pufall博士研究了信号整合 利用糖皮质激素受体（GR），特别是DNA变构调节的机制基础， 顺序这个K99/R 00奖项将为他提供资源、时间和培训来开发系统 将GR变构整合的结构和生物物理学基础转化为对 导致儿童前B急性淋巴细胞白血病（B-ALL）治疗的信号异常 阻力这项工作将使选择性干预的发展，直接变构网络在 蛋白该奖项将使Pufall博士实现以下职业发展目标：1）培养一个 造血发育和疾病的基础; 2）学习与初级组织合作; 3）获得 基因组数据集的经验，以开发未来的研究;和4）开发体外系统，以测试信号 融合原则。 糖皮质激素受体通过以下方式协调响应细胞信号的基因表达程序： 使整个基因组中特定DNA反应元件处的调节复合物的组装成核。 在B-ALL中，针对GR的合成糖皮质激素是一种有效的治疗方法， 其他的药物，可以诱导细胞凋亡。然而，那些对糖皮质激素没有反应的人 面临严峻的预后。Pufall博士假设，变构调节GR的信号通路具有 被破坏，改变受体的正常功能，并阻断细胞凋亡。他会测试这个 假设有三个目的：1）鉴定糖皮质激素诱导的基因表达程序，GR结合， 糖皮质激素敏感和耐药的急性淋巴细胞白血病中活性染色质的标记; 2） 从选择的响应元件纯化GR调节复合物并鉴定组分;和 从GR的DNA结合域发出的变构丝，并确定信号如何冲击这些 电线.

Project Summary/Abstract Proteins lie at the nodes of signaling pathways, and it is their task to integrate these signals to direct a specific output - one that is tailored to the needs of the cell at the time. Dr. Miles Pufall has studied signal integration using the glucocorticoid receptor (GR), in particular the mechanistic basis of allosteric regulation by DNA sequence. This K99/R00 award will provide him with the resources, time, and training to develop systems that translate the structural and biophysical basis of GR allosteric integration into an understanding of the signaling aberrations that result in childhood pre-B acute lymphoblastic leukemia (B-ALL) treatment resistance. This work will enable development of selective interventions that direct allosteric networks in the protein. This award will allow Dr. Pufall to achieve the following career development goals: 1) Develop a grounding in hematopoietic development and disease; 2) Learn to work with primary tissue; 3) Gain experience with genomic data sets to develop future studies; and 4) Develop an in vitro system to test signal integration principles. The glucocorticoid receptor orchestrates a program of gene expression in response to cellular signals by nucleating the assembly of regulatory complexes at specific DNA response elements throughout the genome. In B-ALL, synthetic glucocorticoids directed against GR are an effective treatment, which in combination with other drugs, work to induce an apoptotic program. However those who do not respond to glucocorticoids face a grim prognosis. Dr. Pufall hypothesizes that signaling pathways that allosterically regulate GR have been disrupted, changing the normal function of the receptor, and blocking apoptosis. He will test this hypothesis in three aims: 1) Identify changes in glucocorticoid induced gene expression programs, GR binding, and marks for active chromatin in glucocorticoid sensitive and resistant acute lymphoblastic leukemia; 2) Purify GR regulatory complexes from select response elements and identify components; and 3) Identify allosteric wires emanating from the DNA binding domain of GR, and determine how signals impinge on these wires.