Regulation of methionine metabolism in Bacillus subtilis

枯草芽孢杆菌蛋氨酸代谢的调控

基本信息

批准号：
8450161
负责人：
TINA M. HENKIN
金额：
$ 31.1万
依托单位：
OHIO STATE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2001
资助国家：
美国
起止时间：
2001-08-01 至 2016-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8450161
关键词：
Affect Affinity Animal Model Archaea Bacillus subtilis Bacteria Binding Biochemical Biological Process Boxing Cells Code Complex DNA Sequence Rearrangement Elements Engineering Eukaryota Family Gene Expression Gene Expression Regulation Genes Genetic Transcription Goals Growth Hybrids In Vitro Individual Investigation Laboratories Ligand Binding Ligands Lysine Mediating Metabolic Methionine Metabolism Pathway Molecular Monitor Organism Pathogenicity Physiological Positioning Attribute Property RNA RNA Binding Regulation Regulatory Element Research Role S-Adenosylmethionine Signal Transduction Specificity Structure System Temperature Testing Thiamine Pyrophosphate Trans-Activators Transcript Translation Initiation Variant Virulence Work analog attenuation base cis acting element in vivo insight member methionine adenosyltransferase mutant novel pathogen public health relevance response tool transcription termination

项目摘要

DESCRIPTION (provided by applicant): Riboswitch RNAs represent an important regulatory mechanism in bacteria. RNAs of this type consist of complex elements positioned in the leader region of a transcript, upstream of the regulated coding sequence(s). These RNA elements directly sense a physiological signal that induces a structural change in the RNA, resulting in an effect on downstream gene expression. This project will focus primarily on two classes of S-adenosylmethionine (SAM)-binding riboswitch RNAs, the S box and the SMK box, with additional efforts on the lysine-binding L box, the thiamine pyrophosphate-binding Thi box, and RNA thermosensors that respond to changes in temperature. The first major goal of the project is to investigate the molecular basis for specific ligand recognition by multiple riboswitch RNAs, and the features responsible for differential SAM sensitivity in natural variants of the S box riboswitch. These efforts will also include using the information obtained to engineer novel classes of riboswitches with the goal of testing the predictive power of these analyses. The second major goal is to investigate the structural and functional differences between riboswitches that operate at the transcriptional and translational levels, to test the hypothesis that translational riboswitches (like the SMK box) have the potential to operate reversibly in vivo, allowing multiple regulatory decisions within the lifetime of a single RNA transcript. These studies will include detailed analysis of the ligand-free form of the SMK box RNA and the transition between the ligand-free and ligand-bound forms. The third major goal involves analysis of the interplay between riboswitch elements and other regulatory mechanisms, using the Bacillus subtilis metK gene, encoding SAM synthetase, as an example. Overall, this project will provide basic information about novel RNA-based mechanisms of gene regulation, and will also provide insight into metabolic regulation in pathogenic organisms that use these mechanisms. Gram- positive pathogens generally use regulatory mechanisms closely related to those found in Bacillus subtilis, the model organism for this work. Expression of determinants for pathogenicity are often regulated in response to physiological signals, and understanding how the cell monitors these signals is important for understanding bacterial virulence. It is also likely that many new riboswitch-like mechanisms remain to be uncovered, and the proposed work will provide important tools for investigation of these mechanisms, and predicting how they function within the cell.

描述（由申请人提供）：核糖开关RNA代表细菌中重要的调节机制。这种类型的RNA由位于转录本的领导区域的复杂元素组成，该元素是调节编码序列的上游。这些RNA元素直接感知了诱导RNA的结构变化的生理信号，从而对下游基因表达产生影响。该项目将主要关注两类的S-腺苷甲硫代氨酸（SAM）结合的核糖开关RNA，S盒和SMK盒，并在赖氨酸结合L盒中进行了额外的努力，Thiamine thiamine pyrophophathate-thi thi盒和RNA热盒以及对温度变化响应的RNA热测量器。该项目的第一个主要目标是研究多个核糖开关RNA的特定配体识别的分子基础，以及负责S Box Riboswitch自然变体中差异SAM灵敏度的特征。这些努力还将包括使用为工程师新颖的核糖开关类别获得的信息，以测试这些分析的预测能力。第二个主要目标是研究在转录和翻译水平上运行的核糖开关之间的结构和功能差异，以检验以下假设：转化核糖开关（如SMK框）有可能在体内可逆地操作，从而可以在单个RNA转录的一生内进行多个调节性决策。这些研究将包括对SMK盒RNA的无配体形式的详细分析以及无配体和配体结合形式之间的过渡。第三个主要目标涉及对核糖开关元件与其他调节机制之间的相互作用进行分析，该机制使用枯草芽孢杆菌METK基因，用编码SAM合成酶进行了示例。总体而言，该项目将提供有关基于基因调节的新型机制的基本信息，还将提供对使用这些机制的致病生物代谢调节的见解。革兰氏阳性病原体通常使用与枯草芽孢杆菌（该作品模型生物体）中发现的调节机制。通常根据生理信号来调节病原性决定因素的表达，并了解细胞监测这些信号对于理解细菌毒力的重要性。许多新的类似核糖开关的机制也可能有待发现，拟议的工作将为研究这些机制提供重要的工具，并预测它们在细胞中的功能。