Regulatory Role of Tandem Tryptophan Codons in Chlamydial Persistence

串联色氨酸密码子在衣原体持久性中的调节作用

• 批准号: 10077111
• 负责人: REY A CARABEO
• 金额: $ 59.83万
• 依托单位: UNIVERSITY OF NEBRASKA MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-02-08 至 2024-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10077111
• 关键词: Abnormal Cell; Address; Affect; Amino Acids; Anabolism; Antibiotic Resistance; Antigens; Antimicrobial Resistance; Bacteria; Biological; Biological Assay; Blindness; CRISPR interference; Cell division; Cells; ChIP-seq; Chlamydia; Chlamydia Infections; Chronic; Codon Nucleotides; Communication; Critical Pathways; Development; Disease; Environment; Genes; Genetic Transcription; Genome; Goals; Growth; Healthcare Systems; Human; Immune response; Immunologics; Indoles; Infectious Agent; Infertility; Inflammatory Response; Interferons; Internet; Iron; Label; Lead; Link; Location; Mediating; Medical; Membrane Proteins; Messenger RNA; Metabolic; Modeling; Molecular; Molecular Chaperones; Monitor; Morbidity - disease rate; Morphology; Nature; Nutrient; Nutritional; Organelles; Organism; Pathway interactions; Patients; Pelvic Inflammatory Disease; Pharmacology; Phenotype; Physiological; Pneumonia; Positioning Attribute; Process; Protein Biosynthesis; Protein Precursors; Proteins; Proteomics; Regulation; Regulatory Element; Regulon; Reiter Disease; Respiratory Tract Infections; Ribosomes; Rickettsia; Role; Sampling; Sexually Transmitted Diseases; Starvation; Stress; Suggestion; Symptoms; System; Testing; Therapeutic Intervention; Time; Toxin; Trachoma; Transcript; Transcriptional Regulation; Translating; Translations; Tryptophan; Type III Secretion System Pathway; Vesicle; Work; antitoxin; genetic approach; human pathogen; infection burden; knock-down; new therapeutic target; novel diagnostics; novel strategies; pathogen; pathogenic bacteria; programs; response; stem; therapeutic candidate; transcriptome; transcriptome sequencing; tubal infertility

Project Summary: Regulatory Role of Tandem Trp Codons in Chlamydial Persistence Chlamydia is an obligate intracellular bacterial pathogen that causes a range of serious diseases in humans. It is the primary cause of bacterial sexually transmitted infections (STI) and the causative agent of infectious preventable blindness, trachoma. The major concern of chlamydial infections is that they are often asymptomatic and undiagnosed, which can lead to chronic sequelae, such as pelvic inflammatory disease and tubal factor infertility. Consequently, chlamydial diseases remain a significant burden on health care systems around the world. In adapting to obligate intracellular growth, Chlamydia has significantly reduced its genome size and eliminated genes from various pathways as it relies on the host cell for its metabolic needs. Most bacteria respond to amino acid limitation by engaging a stringent response, which is a transcriptional program used to adapt to nutrient-poor conditions. The stringent response has also been directly connected with inducing persister cells in various types of bacteria, which have limited metabolic activity and increased antimicrobial resistance. Chlamydia can become persistent when limited for nutrients such as tryptophan (trp). They are morphologically aberrant, display a dysregulated transcriptome, non-replicating, yet remain viable. Chlamydia lacks the genes necessary to deploy a stringent response, and has very few identified regulatory elements. How it responds to stress is an intriguing question. We hypothesize that during trp limitation the decreased translation of key Chlamydia genes with tandem trp codons triggers a persistent phenotype. The first project goal is to determine how trp limitation impacts the transcriptional regulation by the iron-dependent YtgR repressor. We will use a combination of next-gen RNA sequencing, including ChIP-seq, and targeted transcriptional assays to address this. The second project goal is to determine the mechanism of chlamydial cell division blockage during persistence by focusing on several cell division genes containing tandem trp codons. A combination of genetic approaches and morphological studies will be used to determine this. The third project goal is to monitor host-pathogen interactions, including nutrient acquisition, via cell biological studies. Because several genes associated with host-pathogen interactions contain tandem trp codons, how the inclusion interacts with host pathways and organelles is predicted to be altered by trp starvation; pathways involved in inclusion integrity are maintained whereas those that support chlamydial progression through the developmental cycle are negatively impacted. The proposal aims to mechanistically link trp starvation, regulation of translation by tandem trp codon motifs, and physiological abnormalities linked to persistence. Results will lead to the identification of novel diagnostic and therapeutic targets that may identify and treat asymptomatic chlamydial infections.

项目摘要：串联TRP密码子在衣原体持久性中的监管作用 衣原体是一种义务的细胞内细菌病原体，会导致一系列严重疾病 人类。它是细菌性传播感染（STI）和因果关系的主要原因 感染性可预防失明的药物，沙眼瘤。衣原体感染的主要关注点是 它们通常是无症状的且未诊断的，可能导致慢性后遗症，例如骨盆 炎症性疾病和输卵管因子不育症。因此，衣原体疾病仍然是重要的 全球医疗保健系统的负担。为了适应义务细胞内生长， 衣原体大大降低了其基因组大小，并从各种途径中消除了基因 依靠宿主细胞的代谢需求。大多数细菌应对氨基酸的限制。 参与严格的响应，这是用于适应营养贫困的转录程序 状况。严格的响应也已与诱导持久细胞中的细胞直接连接 各种类型的细菌，它们的代谢活性有限，抗菌素耐药性增加。 当限制诸如色氨酸（TRP）之类的营养时，衣原体可能会变得持久。他们是 在形态上异常，表现出失调的转录组，不复制，但仍然可行。 衣原体缺乏部署严格反应所需的基因，很少有人确定 监管要素。它如何应对压力是一个有趣的问题。我们假设在TRP期间 限制与串联TRP密码子的关键衣原体基因的翻译减少触发A 持续的表型。第一个项目目标是确定TRP限制如何影响转录 铁依赖性YTGR阻遏物调节。我们将结合下一代RNA 测序，包括ChIP-Seq和针对性的转录测定，以解决此问题。第二个 项目目标是确定在持久性期间的衣原体细胞分裂阻塞的机制 专注于几个包含串联TRP密码子的细胞分裂基因。遗传的结合 方法和形态学研究将用于确定这一点。第三个项目目标是 通过细胞生物学研究，监测宿主 - 病原体相互作用，包括营养的获取。因为 与宿主病原体相互作用相关的几个基因包含串联TRP密码子，如何 纳入与宿主途径相互作用，预计细胞器会因TRP饥饿而改变。 维持包容性完整性涉及的途径，而那些支持衣原体的途径 通过发育周期的发展受到负面影响。该提议旨在 机械上的TRP饥饿，串联TRP密码子图案的翻译调节，以及 与持久性有关的生理异常。结果将导致新颖的识别 可以识别和治疗无症状衣原体感染的诊断和治疗靶标。