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Enabling High-Throughput Analysis and Single-Cell Imaging of Bacterial Signals

实现细菌信号的高通量分析和单细胞成像

基本信息

批准号：
10709561
负责人：
Ming Chen Hammond
金额：
$ 35.57万
依托单位：
UNIVERSITY OF UTAH
依托单位国家：
美国
项目类别：
财政年份：
2017
资助国家：
美国
起止时间：
2017-09-01 至 2026-07-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10709561
关键词：
Affect Animal Model Antibiotic Resistance Antibiotic Therapy Antibiotics Bacteria Behavior Binding Bioinformatics Biological Assay Biology Biosensor Cell Culture Techniques Cell physiology Cells Charge Chemicals Cholera Clinical Combating Antibiotic Resistant Bacteria Complex Cytometry Development Diet Drug Compounding Drug Design Dyes Electron Microscopy Environment Environmental Risk Factor Enzymes Flow Cytometry Fluorescent Dyes Food Poisoning Foundations Funding Gene Expression Genetic Goals Gram-Negative Bacteria Growth Health Health Status Human Image Inflammatory Bowel Diseases Light Link Mammalian Cell Measures Messenger RNA Methods Microbial Biofilms Microbiology Modeling Molecular Genetics Molecular Structure Monitor Noise Nutrient Organism Outcome Pathway interactions Periodicity Permeability Phase Physiological Process Production Productivity Property RNA Regulation Regulator Genes Regulatory Pathway Research Signal Transduction Signaling Molecule Speed Structure Structure-Activity Relationship Surface Technology Time Toxin Typhoid Fever Urinary tract infection Visualization cell motility cellular imaging enzyme activity genetic analysis gut colonization gut health gut microbiota high throughput analysis high throughput screening improved in situ imaging insight invention molecular imaging next generation novel pathogen pathogenic bacteria prebiotics programs public health relevance ratiometric real time monitoring screening small molecule tool uptake

项目摘要

PROJECT SUMMARY Enabling High-Throughput Analysis and Single-Cell Imaging of Bacterial Signals Our research aims to understand how bacteria perceive chemical signals to regulate different behaviors. We have invented different types of biosensors to rapidly measure key signaling molecules in bacteria, including one for cyclic di-GMP. This signal controls whether bacteria attach to surfaces, form sticky biofilms, and secrete toxins. One of our major goals is to identify nutrients, other chemicals, and environmental inputs that change cyclic di-GMP levels in different bacteria. We recently demonstrated a successful approach that combines structure-based bioinformatics analysis and experimental screening. However, the discovery of primary inputs remains challenging because each bacterium harbors many cyclic di-GMP signaling enzymes, the signal is transiently produced, highly charged, and low in abundance, and the screening method remains a key bottleneck. Thus, this proposal will develop next-generation fluorescent biosensors to enhance high-throughput, quantitative screening of enzyme activity directly in cells (Aim 2). These biosensors then will be applied to discover primary inputs for a widespread small molecule binding domain associated with cyclic di-GMP and other signaling enzymes (Aim 3). In addition, towards understanding environmental factors that regulate cyclic di-GMP, this proposal will develop a new type of biosensor to perform in situ imaging of cyclic di-GMP in biofilms (Aim 3). In the long term, this project aims to inform personalized diets to treat inflammatory bowel diseases and promote gut health. For this renewal of the project, the original scope also has been expanded to study the permeability of small molecules into bacterial cells. The permeability process includes passive permeation, active uptake, and active efflux mechanisms, and is critical to bacterial growth, signaling, and antibiotic resistance. This proposal will develop a high-throughput assay that enables real-time monitoring of small molecule permeability in cells (Aim 1). The assay will be applied to understand both the molecular structures and genetic factors that affect accumulation of fluorescent dyes and of clinical antibiotics inside cells. In the long term, this new aim will improve chemical biology tools that use these dyes and antibiotics treatments.

项目总结

项目成果

期刊论文数量（13）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Structure and mechanism of a Hypr GGDEF enzyme that activates cGAMP signaling to control extracellular metal respiration

DOI：
10.7554/elife.43959
发表时间：
2019-04-09
期刊：
ELIFE
影响因子：
7.7
作者：
Hallberg, Zachary F.;Chan, Chi Ho;Hammond, Ming C.
通讯作者：
Hammond, Ming C.

Guanidine Biosensors Enable Comparison of Cellular Turn-on Kinetics of Riboswitch-Based Biosensor and Reporter.

DOI：
10.1021/acssynbio.0c00583
发表时间：
2021-03-19
期刊：
ACS synthetic biology
影响因子：
4.7
作者：
Manna S;Truong J;Hammond MC
通讯作者：
Hammond MC

Second messengers and divergent HD‐GYP phosphodiesterases regulate 3′,3′‐cGAMP signaling

DOI：
10.1111/mmi.14412
发表时间：
2020-01
期刊：
Molecular Microbiology
影响因子：
3.6
作者：
Todd A. Wright;Lucy Jiang;James J. Park;W. A. Anderson;Ge Chen;Zachary F. Hallberg;Beiyan Nan;M. C. Hammond
通讯作者：
Todd A. Wright;Lucy Jiang;James J. Park;W. A. Anderson;Ge Chen;Zachary F. Hallberg;Beiyan Nan;M. C. Hammond

Synthetic Biology of Small RNAs and Riboswitches.

小RNA和核糖开关的合成生物学。