Role of the transcriptional intermediary factor TIF1g in vertebrate hematopoiesis

候选人摘要 我的职业目标是领导一个独立的研究小组，研究自然和 控制造血细胞命运的染色质因子的功能。我的研究背景是 表观遗传基因调控加上我在斑马鱼遗传学方面的专业知识， 造血为我提供了进行拟议研究的知识。的 在儿童医院和哈佛的血液学/肿瘤学部门的培训计划 医学院提供了一个出色的环境，在培训期间完成 指导阶段。这将极大地促进我向独立的顺利过渡。 研究描述 造血是由复杂的遗传程序控制的，涉及组织特异性 转录因子和染色质重塑因子。了解监管 造血机制提供了重要的洞察人类的病理生理学 血液恶性肿瘤，如白血病。转录中介因子TIF 1是一个关键的 造血因子，但其机制还不清楚。通过大规模 使用斑马鱼TIF 1 <$突变体进行遗传抑制筛选，我确定了两个抑制基因， 突变体，可以绕过TIF 1 <$$>的要求，并恢复TIF 1 <$-缺陷的血液 动物这些突变体的初步特征表明，TIF 1在以下方面发挥着重要作用： 调节造血过程中的转录延伸和染色质重塑。的 本提案中描述的研究旨在阐明TIF 1？ 规范这些过程。Aim 1将使用染色质免疫沉淀（ChIP）分析， 彻底检查RNA聚合酶II和相关组蛋白标记物在 TIF 1缺陷细胞中的血液基因。Aim 2将使用可用的条件性基因敲除小鼠， 研究TIF 1 <$及其抑制因子在哺乳动物造血中的功能。AIM 3将 专注于深入表征TIF 1 <$和粘附素之间的相互作用， 染色质重塑复合物使用遗传和生物化学方法。完成 这些目标将揭示转录因子、延伸因子和 造血过程中的染色质重塑。考虑到转录因子的参与， 延伸和染色质修饰在各种人类疾病，这些研究将 推进我们对它们在这些疾病的发病机制和进展中的作用的理解， 疾病，也可以确定候选基因或途径，可用于发展 新的靶向治疗策略。

Candidate Summary My career goal is to direct an independent research group studying the nature and function of chromatin factors that control hematopoietic cell fate. My research background in epigenetic gene regulation plus my developing expertise in zebrafish genetics and hematopoiesis provide me with the knowledge to perform the proposed research. The training program in the hematology/oncology division at Children's Hospital and Harvard Medical School provides an outstanding environment for the completion of training during the mentored phase. This will greatly facilitate my smooth transition to independence. Research Description Hematopoiesis is controlled by complicated genetic programs involving tissue-specific transcription factors and chromatin remodeling factors. Understanding the regulatory mechanism of hematopoiesis provides significant insight into the pathophysiology of human blood malignancies such as leukemia. The transcription intermediary factor TIF1¿ is a critical factor for hematopoiesis yet the mechanism is not well understood. Through a large-scale genetic suppressor screen using the zebrafish TIF1¿ mutant, I identified two suppressor mutants that can bypass the requirement of TIF1¿ and restore blood in TIF1¿-deficient animals. Initial characterizations of these mutants suggest a fundamental role of TIF1¿ in regulating transcriptional elongation and chromatin remodeling during hematopoiesis. The research described in this proposal is designed to elucidate the mechanism by which TIF1¿ regulates these processes. Aim1 will use chromatin immunoprecipitation (ChIP) analyses to thoroughly examine the distribution of RNA polymerase II and associated histone markers on blood genes in TIF1¿-deficient cells. Aim2 will use the available conditional knockout mice to investigate the function of TIF1¿ and its suppressors in mammalian hematopoiesis. Aim3 will focus on the in-depth characterization of the interaction between TIF1¿ and the cohesin chromatin remodeling complex using genetic and biochemical approaches. Completion of these aims will reveal the interplay among transcription factors, elongation factors and chromatin remodelers during hematopoiesis. Given the involvement of transcriptional elongation and chromatin modification in a variety of human disorders, these studies will advance our understanding of their roles in the pathogenesis and progression of these maladies and may also identify candidate genes or pathways that can be used for developing novel targeted treatment strategies.