Dissection of cis-regulatory Regions in Plants Using Recombination, Allelic Expression, and Site-directed Mutagenesis

利用重组、等位基因表达和定点诱变剖析植物中的顺式调控区域

• 批准号: 1157466
• 负责人: Jennifer Hawkins
• 金额: $ 75.59万
• 依托单位: West Virginia University Research Corporation
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-08-01 至 2017-07-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1157466&HistoricalAwards=false
• 关键词: Dissection; cis; regulatory; Regions; Plants

Intellectual Merit. Central to the understanding of organismal development are the underlying genetic mechanisms that drive variation in gene expression. It is generally agreed that sequence variation in the regulatory regions of genes plays a major role in expression divergence and organismal diversity, yet little is know regarding the structure and function of these complex promoter regions. This project takes a step toward resolving the genesis of this expression variation by dissecting the functional modules of regulatory regions into their controlling units. Evidence from a wide range of eukaryotic organisms indicates that, within individual species, variation in allelic expression is a common phenomenon and occurs at a significantly high frequency in various tissues and during different developmental stages. Using two resource-rich model plant taxa, Arabidopsis thaliana and Zea mays (maize), variation in allelic expression, combined with naturally occurring meiotic recombination, will be used to map the regulatory components of these expression differences for several genes in various plant tissues. The data are expected to reveal the regulatory components of allelic expression variation in two distinct plant lineages and contribute significantly to a more complete understanding of the structure and function of plant promoters. Additionally, other regulatory components of gene expression that do not contribute to allelic expression differences, but do function as cis-regulatory controls, are expected to be discovered using in situ site-directed mutagenesis in upstream regions of the same genes in Arabidopsis. These data, combined with those from the allelic expression portion of the project, will provide a detailed characterization of plant promoter structure and function in the Arabidopsis and maize regions of interest. The completion of this project will contribute to a broader understanding of promoter evolution in plants, specifically with respect to the role of module swapping via meitoic recombination, and will also provide a promising methodology for elucidating mechanisms for the cis-regulatory control of plant gene expression. This project promises not only intellectual advancement in the fields of molecular and evolutionary genetics, but also the development of a number of possible practical industrial technologies designed to enhance and advance agricultural practices.Broader Impacts. This research lends itself to multiple activities involving outreach, the inclusion of members of underrepresented groups, and scientific training of students at all levels. In the state of West Virginia, a small fraction of high school students attend college (~16%), and 20% of those incoming undergraduates are first-generation college students. These projects will provide multiple educational opportunities for these underrepresented students via scientific training in laboratory practices that lead to landmark discoveries, and ultimately to the implementation of innovative real world agricultural applications that benefit society as a whole. Other underrepresented groups will be provided opportunities at WVU via the McNair Scholarship, designed to provide support for students from disadvantaged backgrounds, and the NSF Research Education for Undergraduates (REU) program. Also, a number of educational outreach opportunities exist at WVU via the program for Women in Science and Engineering (WiSE), and the Summer Undergraduate Research Experience (SURE) program. Students will be provided the opportunity to disseminate the results of their work among the broader scientific community at annual international meetings. In addition, this research is designed as part of the WVU Computational Biology Initiative, an interdepartmental effort among the Departments of Biology, Mathematics and Statistics to create undergraduate and graduate programs in the emerging field of bioinformatics. All scripts and data generated will be used to develop new courses under the Initiative.

智力优势。 理解生物体发育的核心是驱动基因表达变异的潜在遗传机制。 人们普遍认为，基因调控区的序列变异在表达差异和生物多样性中起着重要作用，但对这些复杂启动子区的结构和功能知之甚少。该项目通过将调节区的功能模块解剖成它们的控制单元，朝着解决这种表达变异的起源迈出了一步。 来自广泛的真核生物的证据表明，在个体物种内，等位基因表达的变化是一种常见现象，并且在各种组织和不同发育阶段中以显著高的频率发生。 使用两个资源丰富的模式植物类群，拟南芥和玉米（玉米），等位基因表达的变化，结合自然发生的减数分裂重组，将被用来映射这些表达差异的调控成分，在各种植物组织中的几个基因。 这些数据有望揭示两种不同植物谱系中等位基因表达变异的调控成分，并有助于更全面地了解植物启动子的结构和功能。 此外，基因表达的其他调控成分，不有助于等位基因的表达差异，但作为顺式调控控制的功能，预计将被发现在拟南芥中相同基因的上游区域使用原位定点诱变。 这些数据，结合那些从该项目的等位基因表达部分，将提供一个详细的表征植物启动子的结构和功能在拟南芥和玉米地区的利益。 该项目的完成将有助于更广泛地了解植物中启动子的进化，特别是关于通过meitoic重组的模块交换的作用，也将提供一个有前途的方法来阐明植物基因表达的顺式调控控制机制。该项目不仅有望在分子遗传学和进化遗传学领域取得知识进步，而且还有望开发出一些可能的实用工业技术，以加强和促进农业实践。 这项研究有助于开展多种活动，包括外联、吸收代表性不足群体的成员以及对各级学生进行科学培训。 在西弗吉尼亚州，只有一小部分高中生上大学（约16%），20%的新生是第一代大学生。 这些项目将为这些代表性不足的学生提供多种教育机会，通过实验室实践的科学培训，导致里程碑式的发现，并最终实施创新的真实的世界农业应用，使整个社会受益。 其他代表性不足的群体将通过麦克奈尔奖学金在西弗吉尼亚大学提供机会，旨在为来自弱势背景的学生提供支持，以及NSF本科生研究教育（REU）计划。 此外，一些教育推广机会存在于西弗吉尼亚大学通过妇女在科学和工程（WiSE）计划，和夏季本科生研究经验（SURE）计划。 学生将有机会在年度国际会议上向更广泛的科学界传播他们的工作成果。此外，这项研究是作为WVU计算生物学计划的一部分，该计划是生物学，数学和统计学部门之间的跨部门努力，旨在创建生物信息学新兴领域的本科和研究生课程。 所生成的所有脚本和数据将用于在该举措下开发新课程。