Quantification and Characterisation of the DNA phosphorothioate modification in the gut microbiome

肠道微生物组中 DNA 硫代磷酸酯修饰的定量和表征

• 批准号: 248597060
• 负责人: Professorin Dr. Stefanie Kaiser
• 金额: --
• 依托单位: Institut für Pharmazeutische Chemie
• 依托单位国家: 德国
• 项目类别: Research Fellowships
• 财政年份: 2013
• 资助国家: 德国
• 起止时间: 2012-12-31 至 2015-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/248597060?language=en
• 关键词: Quantification; Characterisation; DNA; phosphorothioate; modification

Nucleic acids, like DNA and RNA, are the basis of all life. In addition to the known four letter code of DNA, there are several more natural modifications of the canonical nucleosides. Only recently, the first modification of the DNA backbone was discovered, the phosphorothioate modification. This modification is only formed in a few bacteria. Over the last years, research on the connection of human health and the natural bacteria that inhabit the human body has been in an increased focus of attention. However, this research has not yet analyzed the possible influence of the phosphorothioate modification on the gut microbiome. It is my goal to study the influence of thus modified bacteria on the gut microbiome and its possible role in diseases like chronic inflammatory bowel disease. During my 2 years of research with the Dedon group it will be my goal to quantify phosphorothioate modifications in bacterial communities of the mouse gut microbiome. I will establish a sensitive LC-MS/MS method for analysis of the global phosphorothioate modification level. This method will be used to observe changes in the phosphorothioate modification level in antibiotic treated and control mice. An increase in the phosphorothioate levels would be a first hint for an advantage of bacteria, producing the modification, confronted with antibiotic-induced oxidative stress. To study the possible shifts in bacterial populations upon antibiotic treatment, deep sequencing will be applied. Therefore, enrichment of phosphorothioate modified DNA will be optimized by Hg-affinity purification. The final goal is the comparison of the global phosphorothioate modification level (by LC-MS/MS) and the changes of bacterial populations (deep sequencing) induced by antibiotic treatment. The comparison of the data received from both analytical methods will allow an insight into the natural function of the phosphorothioate modification and its connection to health and disease.

核酸，像DNA和RNA一样，是所有生命的基础。除了已知的DNA的四个字母代码之外，还有几种典型核苷的天然修饰。直到最近，DNA骨架的第一个修饰才被发现，即硫代磷酸酯修饰。这种修饰只在少数细菌中形成。在过去的几年里，关于人类健康与居住在人体内的天然细菌之间的联系的研究一直受到越来越多的关注。然而，这项研究尚未分析硫代磷酸酯修饰对肠道微生物组的可能影响。我的目标是研究这样修饰的细菌对肠道微生物组的影响及其在慢性炎症性肠病等疾病中的可能作用。在我与Dedon小组的两年研究期间，我的目标是量化小鼠肠道微生物组细菌群落中的硫代磷酸酯修饰。我将建立一种灵敏的LC-MS/MS方法，用于分析整体硫代磷酸酯修饰水平。该方法将用于观察抗生素处理小鼠和对照小鼠中硫代磷酸酯修饰水平的变化。硫代磷酸酯水平的增加将是细菌优势的第一个暗示，产生修饰，面临着干旱诱导的氧化应激。为了研究抗生素治疗后细菌种群的可能变化，将应用深度测序。因此，硫代磷酸酯修饰的DNA的富集将通过Hg亲和纯化来优化。最终目标是比较整体硫代磷酸酯修饰水平（通过LC-MS/MS）和抗生素治疗诱导的细菌种群变化（深度测序）。从两种分析方法中获得的数据的比较将允许洞察硫代磷酸酯修饰的天然功能及其与健康和疾病的联系。

项目成果

Novel genomic island modifies DNA with 7-deazaguanine derivatives

• DOI: 10.1073/pnas.1518570113
• 发表时间: 2016-03-15
• 期刊: PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA
• 影响因子: 11.1
• 作者: Thiaville, Jennifer J.;Kellner, Stefanie M.;de Crecy-Lagard, Valerie
• 通讯作者: de Crecy-Lagard, Valerie