Defining New Functions in DNA Replication and DNA Damage Response Genes

定义 DNA 复制和 DNA 损伤反应基因的新功能

• 批准号: 1158560
• 负责人: Victoria Lundblad
• 金额: $ 30万
• 依托单位: The Salk Institute for Biological Studies
• 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: 2012-05-01 至 2014-04-30
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1158560&HistoricalAwards=false
• 关键词: Defining; New; Functions; DNA; Replication

Scientific goals: In the budding yeast S. cerevisiae, genome-wide collections of mutant strains have been employed extensively to elucidate biological function on a large scale. The resulting information has been used to place genes in pathways, to identify points of intersection between different pathways and to assign gene function to novel proteins. One flaw of these studies stems from the nature of the mutant reagents in these genome-wide collections, where gene function has been inactivated by either deletion or conditional depletion. Since many, if not all, proteins execute more than one function, such mutations are potentially pleiotropic. A potential solution is to employ separation-of-function mutations that eliminate a single biological function of a protein. However, identification of this particular class of alleles in the past has been a logistic hurdle even for single genes. This research project is based on a newly developed strategy for large-scale isolation of separation-of-function alleles which will be applied to a set of inter-related genes involved in DNA replication and response to DNA damage. Since very few mutations have been isolated for many of these genes, this research should define new functions in the DNA replication and DNA damage response pathways. Broader impacts: The scientific goals of this research project are closely intertwined with two broader impacts. First, the resulting mutations will be made widely available to the yeast community, with a particular focus on incorporation of these new reagents into genome-wide systems analysis. Genetic networks constructed from separation-of-function missense mutations are likely to uncover previously unappreciated interfaces that were missed in prior systems analysis which employed currently available mutant strains. Second, this project will rely heavily on entry-level researchers (undergraduate and high school students) who will be responsible for generating the panels of separation-of-function mutations. This experience will allow very junior researchers to play a central role in a significant research project and also introduce them to the critical function that mentoring plays in the biomedical research community.

科学目标：在芽殖酵母酿酒酵母中，突变菌株的全基因组集合已被广泛用于大规模阐明生物学功能。由此产生的信息已被用来将基因放置在通路中，识别不同通路之间的交叉点，并将基因功能分配给新的蛋白质。这些研究的一个缺陷源于这些全基因组集合中突变试剂的性质，其中基因功能已因缺失或条件缺失而失活。由于许多（如果不是全部）蛋白质执行不止一种功能，因此此类突变可能是多效性的。一个潜在的解决方案是采用功能分离突变来消除蛋白质的单一生物学功能。然而，在过去，即使对于单个基因来说，鉴定这类特殊的等位基因也是一个后勤障碍。该研究项目基于一项新开发的大规模分离功能分离等位基因的策略，该策略将应用于一组参与 DNA 复制和 DNA 损伤反应的相互关联的基因。由于这些基因中的许多基因分离出的突变很少，因此这项研究应该定义 DNA 复制和 DNA 损伤反应途径中的新功能。更广泛的影响：该研究项目的科学目标与两个更广泛的影响密切相关。首先，由此产生的突变将广泛提供给酵母界，特别关注将这些新试剂纳入全基因组系统分析。由功能分离错义突变构建的遗传网络可能会揭示以前未受重视的界面，这些界面在之前使用当前可用突变株的系统分析中被遗漏了。其次，该项目将严重依赖入门级研究人员（本科生和高中生），他们将负责生成功能分离突变组。这种经验将使初级研究人员能够在重大研究项目中发挥核心作用，并向他们介绍指导在生物医学研究社区中发挥的关键作用。