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

智力价值：这个项目解决了生物学中的一个重要的基本问题。是否存在组蛋白修饰的生物密码来调节基因表达模式？组蛋白是一种与DNA结合形成染色体的小蛋白，组蛋白上的个别位点特异性翻译后修饰与基因激活或沉默有关。然而，组蛋白修饰的同时组合对基因活性的影响尚不清楚。缺乏这方面知识的主要原因是，目前还不存在通过任何标准的生物技术对组蛋白组合修饰进行定量表征甚至定性鉴定的方法。为此，本研究将开发基于蛋白质组学的新方法和伴随的生物信息学，以定量表征组合的组蛋白修饰模式及其在不同生物条件下对基因表达的影响。该项目开发的整合和变革性蛋白质组学技术将立即为广泛的科学家所用。这些实验的结果将在普林斯顿大学Lewis-Sigler整合基因组研究所托管的基于网络的数据库中公开可用。这些研究与生化实验相结合，将有助于提供关于基因表达的系统生物学前景，这将为推进几个应用奠定基本的科学基础，如细胞重新编程。更广泛的影响：这个项目将为社区大学的教育者和学生(包括少数族裔)提供基因组学和蛋白质组学的介绍。尽管有许多现有的计划来支持大学一级的本科生和研究生研究，并向K-12学生推广科学，但针对社区学院学生的这样的计划却很少。将提供多个机会。与当地产业合作伙伴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。暑期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现代蛋白质组学研究的理论和实践。该讲习班将提供课堂讲授和实验室培训，使与会者能够准备样本、获取和解释数据以及学习采集后数据处理。一个包含教程、评论和其他资源的网站将免费提供，作为社区大学教育工作者返回校园后不断更新他们的蛋白质组学培训的外部手段。外展活动将包括通过参加少数群体研究会议，如奇卡诺人和美国原住民科学促进会，以及通过与其他大学的多样性计划(即麦克奈尔学者)直接互动，招募代表人数不足的学生。该项目由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).