TRANSCRIPTION ASSOCIATED MUTAGENESIS IN CELLS UNDER CONDITIONS OF ARRESTED GROWT

生长抑制条件下细胞转录相关的诱变

• 批准号: 8168232
• 负责人: Eduardo A Robleto
• 金额: $ 8.76万
• 依托单位: UNIVERSITY OF NEVADA RENO
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-09-01 至 2011-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8168232
• 关键词: Affect; Amino Acids; Bacillus subtilis; Bacterial Model; Biological Assay; Cell Cycle; Cell Death; Cells; Code; Computer Retrieval of Information on Scientific Projects Database; Degenerative Disorder; Funding; Gene Targeting; Genes; Genetic; Genetic Transcription; Genomic Instability; Grant; Growth; Health; Human; Institution; Lead; Malignant Neoplasms; Mediating; Mutagenesis; Mutation; Organism; Process; Proteins; Research; Research Personnel; Resources; Source; Starvation; Testing; United States National Institutes of Health; cell growth; cell growth regulation; gain of function mutation; novel; pressure; research study; tumor

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. This proposal studies novel mechanisms of mutation that take place in cells under conditions of growth arrest. These mechanisms are of relevance to cell growth and differentiation and human health as they may generate mutations that circumvent growth cell cycle check points, which lead to genome instability, and the formation of tumors or cell death, all processes associated with cancer and other degenerative diseases. The overall hypothesis tested here is that increases in transcription (the making of gene products) of genes under selection in cells in conditions of growth arrest results in accumulation of mutations that promote escape from arrested growth (transcription associated mutagenesis). To examine this hypothesis, we use a bacterial model for cell growth and differentiation in which Bacillus subtilis cells are subjected to amino acid starvation. Under these conditions, cells carrying genetic deficiencies in amino acid synthesis escape arrested growth by acquiring gain of function mutations in the genes coding for synthesis of the limiting amino acid (genes under selective pressure). We ask the question: does altering transcription levels of genes under selection lead to changes in the accumulation of mutations that promote escape from growth arrest? To answer this question, we will use two experimental approaches to assay the effects of modulating transcription of genes under selective pressure: 1) We will genetically inactivate factors mediating transcription and determine whether their inactivation leads to a decrease in expression and accumulation of mutations in a target gene. 2) We will build genetic constructs where expression of target gene can be induced or repressed and assay whether the accumulation of mutations correlates with levels of transcription. Ultimately, the experiments proposed here provide a better understanding of mechanisms of mutation that affect all organisms and are of particular importance to regulation of cell growth and differentiation and to human health.

该子项目是利用该技术的众多研究子项目之一 资源由 NIH/NCRR 资助的中心拨款提供。子项目和 研究者 (PI) 可能已从 NIH 的另一个来源获得主要资金， 因此可以在其他 CRISP 条目中表示。列出的机构是 对于中心来说，它不一定是研究者的机构。 该提案研究了生长停滞条件下细胞中发生的新突变机制。这些机制与细胞生长和分化以及人类健康相关，因为它们可能产生绕过生长细胞周期检查点的突变，从而导致基因组不稳定、肿瘤形成或细胞死亡，所有这些过程都与癌症和其他退行性疾病相关。 。这里测试的总体假设是，在生长停滞条件下细胞中选择的基因转录（基因产物的产生）增加导致突变的积累，从而促进逃避生长停滞（转录相关诱变）。为了检验这一假设，我们使用细胞生长和分化的细菌模型，其中枯草芽孢杆菌细胞遭受氨基酸饥饿。在这些条件下，携带氨基酸合成遗传缺陷的细胞通过在编码限制性氨基酸合成的基因（选择压力下的基因）中获得功能突变来逃避生长停滞。我们提出一个问题：改变选择下基因的转录水平是否会导致突变积累的变化，从而促进逃避生长停滞？为了回答这个问题，我们将使用两种实验方法来测定在选择压力下调节基因转录的效果：1）我们将从基因上灭活介导转录​​的因子，并确定它们的灭活是否会导致基因表达的减少和突变的积累。目标基因。 2）我们将构建可以诱导或抑制靶基因表达的基因构建体，并分析突变的积累是否与转录水平相关。最终，这里提出的实验提供了对影响所有生物体的突变机制的更好理解，并且对于细胞生长和分化的调节以及人类健康特别重要。