RUI: Analysis of the Role of CHD1 in Chromatin Structure and Transcription

染色质重塑因子利用ATP的能量来调节染色质的结构，从而调节诸如转录、复制和修复的过程。 虽然这些ATP酶在体外的作用机制方面已经取得了重大进展，但关于它们在体内的活性仍有许多未知之处。PI的长期目标是了解CHD 1染色质重塑和组装因子如何在体内调节和靶向。Drosophila melanogaster是一个理想的模型系统，在其中研究这种不寻常的protein.Intellectual merit的行动：这项研究的目标是使用果蝇遗传学的强大工具，调查在体内活动的CHD 1，一个chromodomain，解旋酶，和DNA结合域蛋白。CHD 1是一种不寻常的ATP酶，因为它在体外具有核小体重塑和核小体组装活性。 这些实验解决了以下特定假设：CHD 1在全局上发挥作用以在活性基因上重新组装染色质，这是导致转录抑制的活性。这一假设将通过追求三个具体目标来解决：（1）通过检查功能获得和功能丧失的chd 1突变体的表型来研究chd 1的体内功能。(2)确定每个保守蛋白质结构域的相对贡献。(3)检查CHD 1 ATP酶活性的获得和丧失对染色体结构和RNA聚合酶II的影响。PI将采用遗传学和细胞生物学方法的组合来分析突变体表型、染色体形态和转录。 这些成熟的技术适用于本科生研究，并已被PI广泛用于相关染色质重塑因子Brahma和ISWI的研究。PI的初步结果和专业知识使她处于一个独特的位置，以实现这些特定的目标，其结果应提供重要的洞察力的功能和调节CHD 1，一个高度保守但知之甚少ATPase.Broader影响：该研究将主要由本科理科学生从克莱蒙麦肯纳，斯克里普斯和Pitzer学院进行。在她在联合科学系的头三年里，PI亲自在实验室培训了15名本科生（13名女性和2名男性），其中包括两名拉丁裔第一代大学生。学生平均在研究实验室工作两年。在已经毕业的PI研究生中，100%正在研究生院或医学院继续他们的科学教育。在这项研究的三年中，PI将在研究实验室亲自培训另外12-15名本科生，为他们提供技能，帮助他们在科学事业中取得成功。PI位于一个独特的部门（联合科学部），由三所顶级文科院校共享。斯克里普斯学院是一所全女子学院，而克莱蒙麦肯纳学院和皮策学院是顶级文理学院中第5和第7大种族多元化学院。这种多样化的学生群体使PI能够鼓励来自历史上代表性不足的群体的理科学生。除了为本科生提供研究机会外，这项研究还将允许本科生参加除了南加州本科生研究会议之外的年度果蝇研究会议。PI的一名本科研究生是该实验室最新出版物《遗传学》的合著者。RUI的这项研究将为PI和她的学生提供资源，继续他们目前的竞争性研究，利用现代分子遗传技术研究染色质重塑蛋白。

Chromatin remodeling factors use the energy of ATP to modulate the structure of chromatin, thereby regulating processes such as transcription, replication, and repair. While significant advances have been made in regards to the mechanism of action of these ATPases in vitro, much remains unknown regarding their activities in vivo. The PI's long-term goal is to understand how the CHD1 chromatin remodeling and assembly factor is regulated and targeted in vivo. Drosophila melanogaster is an ideal model system in which to study the actions of this unusual protein.Intellectual merit: The goal of this research is to use the powerful tools of Drosophila genetics to investigate the in vivo activities of CHD1, a chromodomain, helicase, and DNA-binding domain protein. CHD1 is an unusual ATPase in that it possesses both nucleosome remodeling and nucleosome assembly activities in vitro. These experiments address the specific hypothesis that CHD1 functions globally to reassemble chromatin on active genes, an activity that results in the inhibition of transcription. This hypothesis will be addressed by pursuing three specific aims (1) Investigate the in vivo function of CHD1 by examining phenotypes of both gain of function and loss of function chd1 mutants. (2) Determine the relative contributions of each of the conserved protein domains. (3) Examine the consequences of both gain and loss of CHD1 ATPase activity on chromosome structure and RNA Polymerase II. The PI will employ a combination of genetic and cell biological approaches to analyze mutant phenotypes, chromosome morphology, and transcription. These well-established techniques are amenable to undergraduate research, and have been extensively used by the PI in her studies of related chromatin remodeling factors Brahma and ISWI. The PI's preliminary result and expertise place her in a unique position to accomplish these specific aims, the results of which should provide significant insights into the function and regulation of CHD1, a highly conserved yet poorly understood ATPase.Broader impacts: The research will be carried out primarily by undergraduate science students from Claremont McKenna, Scripps, and Pitzer Colleges. During her first three years in the Joint Science Department, the PI personally trained 15 undergraduates in the laboratory (13 women and two men), including two Latina first-generation college students. Students spend an average of two years in the research laboratory. Of the PIs research students who have graduated, 100% are continuing their science education in graduate or medical schools. Over the three years of this study, the PI will personally train an additional 12-15 undergraduates in the research laboratory, providing them with skills to help them succeed in their careers in science. The PI is in a unique department (The Joint Science Department) that is shared by three top-tier liberal arts institutions. Scripps is an all-women college, while Claremont McKenna and Pitzer College are the 5th and 7th most ethnically diverse liberal arts colleges in the top tier. This diverse student body allows the PI to encourage science students from historically underrepresented groups. In addition to providing research opportunities for undergraduates, this research will allow undergraduates to attend the Annual Drosophila Research Conference in addition to the Southern California Conference for Undergraduate Research. One of the PI's undergraduate research students is a co-author on the lab's most recent publication in Genetics. This RUI research will provide resources for the PI and her students to continue their current, competitive research into chromatin remodeling proteins using modern molecular genetic techniques.