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: 7959720
• 负责人: Eduardo A Robleto
• 金额: $ 12.88万
• 依托单位: UNIVERSITY OF NEVADA RENO
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-06-01 至 2010-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7959720
• 关键词: Affect; Amino Acids; Bacillus subtilis; Bacterial Model; Biological Assay; Biomedical Research; 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.

该副本是利用众多研究子项目之一 由NIH/NCRR资助的中心赠款提供的资源。子弹和 调查员（PI）可能已经从其他NIH来源获得了主要资金， 因此可以在其他清晰的条目中代表。列出的机构是 对于中心，这不一定是调查员的机构。 该建议研究在生长停滞条件下在细胞中发生的新型突变机制。这些机制与细胞的生长，分化和人类健康有关，因为它们可能会产生突变，从而规避生长细胞周期检查点，从而导致基因组不稳定性以及肿瘤或细胞死亡的形成，所有过程与癌症和其他退化性疾病有关。此处检验的总体假设是，在生长停滞条件下，在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，这些突变促进了促进避免生长的逃脱（转录相关的诱变）。为了审查这一假设，我们使用细菌模型进行细胞生长和分化，其中枯草芽孢杆菌细胞受到氨基酸饥饿。在这些条件下，氨基酸合成中携带遗传缺陷的细胞通过获取编码有限氨基酸（选择性压力下的基因）的基因中的功能突变而避免了生长。我们提出一个问题：选择在选择下的基因的转录水平是否会导致突变积累的变化，这些突变促进了逃避生长停滞的逃脱？为了回答这个问题，我们将使用两种实验方法来测定选择性压力下基因转录的影响：1）我们将在遗传上介导转录的遗传失活因子，并确定它们的失活是否导致靶基因中突变的表达和突变的积累下降。 2）我们将建立遗传构建体，其中可以诱导或抑制靶基因的表达，并测定突变的积累是否与转录水平相关。最终，这里提出的实验可以更好地理解影响所有生物的突变机制，并且对于调节细胞生长和分化以及对人类健康至关重要。