Identifying and functionally characterizing the molecular determinants of chromosome stability in higher eukaryotes

鉴定和功能表征高等真核生物染色体稳定性的分子决定因素

• 批准号: RGPIN-2018-05007
• 负责人: McManus, Kirk
• 金额: $ 7.29万
• 依托单位: University of Manitoba
• 依托单位国家: 加拿大
• 项目类别: Discovery Grants Program - Individual
• 财政年份: 2022
• 资助国家: 加拿大
• 起止时间: 2022-01-01 至 2023-12-31
• 项目状态: 已结题
• 来源: https://www.nserc-crsng.gc.ca/ase-oro/Details-Detailles_eng.asp?id=744196
• 关键词: Identifying; functionally; characterizing; molecular; determinants

A stable genome is a fundamental property of all eukaryotes that is essential for the maintenance and accurate partitioning of DNA into daughter cells. Thus, genome stability is critical for normal cell function, physiology and species survival. Chromosome stability (CS), or the accurate maintenance of normal chromosome numbers and DNA content, is vital for genome stability, yet a huge gap exists in our fundamental understanding of the genes, proteins and cellular processes (i.e. molecular determinants) that regulate CS. My NSERC Discovery Program (DP) will fill this gap by identifying and functionally characterizing the molecular determinants of CS, to provide unprecedented insight into genome stability and normal cell function.My DP has developed novel, quantitative imaging tools to identify and functionally evaluate CS genes and proteins. Bolstered by an industrial partnership that enhances my analytical capabilities and throughput, I am now applying these innovative tools to gain unparalleled insight into the molecular determinants of CS in higher eukaryotes. My initial focus stems from my unpublished pilot work that identified three members of the Condensin Complex as potential CS genes. In humans, there are two Condensin complexes (8 total proteins) that function to compact DNA into chromosomes, particularly during mitosis. Data from others suggest that aberrant chromosome compaction adversely impacts mitotic fidelity and leads to the loss of CS; however, the link between Condensin and CS has not been fully elucidated. Using genetic, biochemical and cell biology methods, I will gain a detailed understanding of the fundamental roles each Condensin protein has in CS. To gain further insight into the molecular determinants of CS, I will screen 501 candidate CS genes identified through cross-species approaches. I will quantitatively assess each gene as above, and generate a prioritized list from which I will functionally evaluate the top 10 candidates to determine their fundamental roles in CS. Collectively, my DP will yield important new knowledge that will advance our fundamental understanding of CS and its impact on genome stability and normal cell function in higher eukaryotes. More specifically, my DP will define the roles each Condensin protein has in CS, and will identify and functionally characterize novel genes and proteins to determine their essential roles in CS. Accordingly, my DP will impact and be of interest to geneticists and cell biologists, but will also generate applied and translational interest as well.Trainees will benefit from my highly productive and collaborative research program. Trainees will receive comprehensive technical, conceptual and theoretical training in genetics, molecular/cell biology and CS to advance their specific projects, which will also strategically position them for related training and job prospects within academia and industry.

稳定的基因组是所有真核生物的基本属性，对于维持和准确地将DNA分配到子细胞中至关重要。因此，基因组稳定性对于正常细胞功能、生理学和物种生存至关重要。染色体稳定性（CS），或正常染色体数目和DNA含量的准确维持，对基因组稳定性至关重要，但我们对调控CS的基因、蛋白质和细胞过程（即分子决定簇）的基本理解存在巨大差距。我的NSERC发现计划（DP）将通过识别和功能表征CS的分子决定因素来填补这一空白，为基因组稳定性和正常细胞功能提供前所未有的见解。我的DP开发了新的定量成像工具来识别和功能评估CS基因和蛋白质。在工业合作伙伴关系的支持下，提高了我的分析能力和通量，我现在正在应用这些创新工具，以获得对高等真核生物中CS分子决定因素的无与伦比的洞察力。我最初的关注源于我未发表的试点工作，该工作确定了冷凝蛋白复合物的三个成员作为潜在的CS基因。在人类中，有两种凝聚蛋白复合物（8种总蛋白质），其功能是将DNA压缩成染色体，特别是在有丝分裂期间。来自其他人的数据表明，异常的染色体压实对有丝分裂保真度产生不利影响，并导致CS的丢失;然而，Condensin和CS之间的联系尚未完全阐明。利用遗传学、生物化学和细胞生物学方法，我将详细了解每个凝聚蛋白在CS中的基本作用。为了进一步了解CS的分子决定因素，我将筛选501个通过跨物种方法确定的候选CS基因。我将定量评估上述每个基因，并生成一个优先级列表，从中我将功能评估前10名候选人，以确定他们在CS中的基本作用。总的来说，我的DP将产生重要的新知识，这将促进我们对CS及其对高等真核生物基因组稳定性和正常细胞功能的影响的基本理解。更具体地说，我的DP将定义每个凝聚蛋白在CS中的作用，并将识别和功能表征新的基因和蛋白质，以确定它们在CS中的重要作用。因此，我的博士论文将影响并引起遗传学家和细胞生物学家的兴趣，但也将产生应用和翻译兴趣。学员将从我的高生产力和合作研究计划中受益。学员将接受遗传学，分子/细胞生物学和CS方面的全面技术，概念和理论培训，以推进他们的具体项目，这也将使他们在学术界和工业界的相关培训和就业前景中处于战略地位。