CAREER: Quantitative Mass Spectrometry Analysis of Histone Codes in Saccharomyces cerevisiae

智力优点：这个项目解决了生物学中一个重要的基本问题。 组蛋白上是否存在一种生物学密码来调节基因的表达模式？ 组蛋白是与DNA结合以形成染色体的小蛋白质，并且组蛋白上的个体位点特异性翻译后修饰与基因激活或沉默相关。 然而，组蛋白修饰的同时组合对基因活性的影响尚不清楚。 这种知识缺乏的主要原因是不存在通过任何标准生物技术对组合组蛋白修饰进行定量表征或甚至定性鉴定的方法。 为此，本研究将开发新的基于质谱的蛋白质组学方法和伴随的生物信息学定量表征组合组蛋白修饰模式及其对酵母中不同生物条件下基因表达的影响。 该项目开发的综合性和变革性蛋白质组学技术将立即被广泛的科学家使用。 这些实验的结果将在普林斯顿大学刘易斯-西格勒综合基因组学研究所托管的基于网络的数据库中公开。 这些研究与生物化学实验相结合，将有助于提供基因表达的系统生物学前景，这将为推进细胞重编程等几种应用奠定基本的科学基础。 更广泛的影响：该项目将向社区学院一级的教育工作者和学生（包括少数民族）介绍基因组学和蛋白质组学。虽然有许多现有的计划，以支持本科生和研究生在大学一级的研究，并促进科学的K-12学生，很少有这样的计划存在的社区学院的学生。 将提供多种机会。 与当地工业合作者建立了战略伙伴关系，以启动支持社区大学生暑期研究经验的机制。 一个夏季蛋白质组学研讨会将用于教育和培训社区大学教育工作者在现代蛋白质组学研究的理论和实践。 该研讨会将提供课堂讲座和实验室培训，使与会者能够准备样品，获取和解释数据，并学习获取后的数据处理。 一个包含教程、评论和其他资源的网站将免费提供，作为社区学院教育工作者返回校园后不断更新蛋白质组学培训的外部手段。 外展计划将包括通过参加少数民族研究会议，如在科学促进奇卡诺斯和美洲原住民的社会，并通过与其他学院的多样性计划（即麦克奈尔学者）的直接互动招聘代表性不足的学生。该项目是由CHE（生命过程化学）和MCB（基因和基因组系统）共同资助。

Intellectual Merit: This project addresses an important fundamental question in biology. Is there a biological code of modifications on histone proteins that regulate gene expression patterns? Histones are small proteins that associate with DNA to form chromosomes, and individual site-specific post-translational modifications on the histones are associated with gene activation or silencing. However, the effect that simultaneous combinations of histone modifications have on gene activity is not known. The main reason for this lack of knowledge is that methods for quantitative characterization, or even qualitative identification, of combinatorial histone modifications by any standard biological technique do not exist. Towards this end, this research will develop novel mass spectrometry based proteomic methods and accompanying bioinformatics to quantitatively characterize combinatorial histone modification patterns and their influence on gene expression under different biological conditions in yeast. The integrated and transformative proteomic techniques developed by this project will be of immediate use to a wide range of scientists. Results of these experiments will be made publically available in a web based database hosted through the Lewis-Sigler Institute for Integrative Genomics at Princeton University. These studies in combination with biochemical experiments will help provide a systems biology outlook on gene expression that will lay the basic scientific foundation to advance several applications, such as cellular reprogramming. Broader Impacts: This project will provide an introduction to genomics and proteomics to both educators and students (including minorities) at the community college level. Although there are many existing programs to support undergraduate and graduate research at the university level and to promote science to K-12 students, few such programs exist for community college students. Multiple opportunities will be made available. Strategic partnerships with local industrial collaborators have been created to initiate a mechanism of support for community college student summer research experiences. A summer proteomics workshop will be used to educate and train community college educators in theory and practice of modern proteomics research. This workshop will offer classroom lectures and laboratory training, allowing the attendees to prepare samples, acquire and interpret data and learn post-acquisition data processing. A website containing tutorials, reviews and other resources will be made freely available as an external means for the community college educators to continuously update their proteomics training upon returning to their campuses. Outreach initiatives will include recruitment of underrepresented students through participation in minority research conferences such as at the Society for the Advancement of Chicanos and Native Americans in Science, and through direct interaction with diversity programs (i.e. McNair Scholars) at other colleges.This project is co-funded by CHE (Chemistry of Life Processes) and MCB (Genes and Genome Systems).