GENETIC INSTABILITY IN YEAST

酵母的遗传不稳定性

• 批准号: 6447959
• 负责人: BONITA J BREWER
• 金额: $ 24.84万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-04-17 至 2002-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6447959
• 关键词: DNA binding protein; DNA replication; Saccharomyces cerevisiae; Werner's syndrome; fungal proteins; gene mutation; helicase; human genetic material tag; molecular pathology; nucleic acid metabolism; protein protein interaction; protein structure function; yeast two hybrid system

DESCRIPTION: (Applicant's Description) In a variety of organisms, mutations that cause depletion of DNA replication enzymes increase the incidence of genetic instability-- events that are characterized by elevated rates of recombination deletion, and/or chromosome loss. Many of the cellular phenotypes of Werner Syndrome (WS) suggest that it falls into this class of genetic defect. The Werner syndrome gene WRN was recently cloned and sequenced and was found to encode a DNA helicase of the RecQ family. The applicants propose that cells that carry out replication in the absence of the WRN helicase leave behind lesions in the DNA that are repaired by a recombination system that produces the genomic rearrangements. They plan to explore this possibility in yeast, by looking for specific types of replication defects that might be generated in the absence of the yeast WRN helicase homologue, Sgs1p. In addition, by studying the types of recombination events induced in yeast cells deficient for the Sgs1 protein, and by looking for proteins that interact with both the normal WRN and Sgs1 helicases, they hope to gain insight into the types of primary damage generated by the replication machinery in the absence of this helicase. The identification of yeast proteins with homology to human proteins and the ability to study them genetically in yeast has led to an understanding of many processes that contribute to human disease. Because of the small genome size of S. cerevisiae-- recently sequenced in its entirety-- the physical analysis of chromosomes during their replication is quite feasible. In no other eukaryotic organism have the cis-acting elements and trans-acting proteins for replication initiation been as well characterized as they have been in yeast. Moreover, the ease with which the genome can be manipulated makes yeast an ideal host organism in which to study basic chromosome biology. Finally, yeast provides a user-friendly cellular environment in which to study protein-protein interactions by way of the two-hybrid transcriptional reporter system.

描述：（申请人的描述）在多种生物体中，导致DNA复制酶耗竭的突变增加了遗传不稳定性的发生率——其特征在于重组缺失和/或染色体丢失率升高。 维尔纳综合征 (WS) 的许多细胞表型表明它属于此类遗传缺陷。 维尔纳综合征基因WRN最近被克隆并测序，并被发现编码RecQ家族的DNA解旋酶。 申请人提出，在没有WRN解旋酶的情况下进行复制的细胞会在DNA中留下损伤，这些损伤可以通过产生基因组重排的重组系统进行修复。 他们计划通过寻找在缺乏酵母 WRN 解旋酶同源物 Sgs1p 的情况下可能产生的特定类型的复制缺陷，来探索酵母中的这种可能性。 此外，通过研究缺乏 Sgs1 蛋白的酵母细胞中诱导的重组事件类型，并寻找与正常 WRN 和 Sgs1 解旋酶相互作用的蛋白，他们希望深入了解 Sgs1 蛋白缺陷产生的主要损伤的类型。在没有该解旋酶的情况下的复制机器。鉴定与人类蛋白质同源的酵母蛋白以及在酵母中对它们进行遗传研究的能力使人们了解了导致人类疾病的许多过程。 由于酿酒酵母的基因组较小（最近已对其进行了整体测序），因此在其复制过程中对染色体进行物理分析是相当可行的。在任何其他真核生物中，用于复制起始的顺式作用元件和反式作用蛋白都没有像在酵母中那样得到如此充分的表征。 此外，基因组操作的简便性使酵母成为研究基本染色体生物学的理想宿主生物。 最后，酵母提供了一个用户友好的细胞环境，在其中通过双杂交转录报告系统研究蛋白质-蛋白质相互作用。