PlrSR-dependent Signal Transduction in Bordetella Virulence

博德特氏菌毒力中 PlrSR 依赖性信号转导

• 批准号: 10097965
• 负责人: Peggy A Cotter
• 金额: $ 49.79万
• 依托单位: UNIV OF NORTH CAROLINA CHAPEL HILL
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-03-06 至 2023-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10097965
• 关键词: Adolescent and Young Adult; Aerobic; Affect; Affinity; Air; Anaerobic Bacteria; Bacteria; Binding; Binding Sites; Biochemical; Biological; Bordetella; Bordetella bronchiseptica; Bordetella pertussis; Carbon Dioxide; Cell Survival; Cells; Cessation of life; ChIP-seq; Child; Country; DNA Sequence; Data; Deoxyribonuclease I; Developed Countries; Developing Countries; Development; Disease; EMSA; Epitopes; Family; Gene Activation; Gene Expression; Gene Expression Profile; Gene Expression Regulation; Genes; Genetic; Genetic Recombination; Heme; Human; Immunity; Immunization; Immunize; In Vitro; Incidence; Individual; Infant; Infection; Light; Link; Lower respiratory tract structure; Mammals; Mediating; Modeling; Molecular; Mus; Nasal cavity; Oxidases; Oxidation-Reduction; Oxides; Oxygen; Pathogenesis; Pertussis; Pertussis Vaccine; Phosphoric Monoester Hydrolases; Phosphotransferases; Population; Protein Family; Proteins; Public Health; Rattus; Regulation; Risk; Role; Signal Transduction; Structural Genes; System; Testing; United States; Vaccinated; Vaccination; Vaccines; Virulence; Virulence Factors; base; cytochrome c oxidase; diagnostic assay; experimental study; genetic approach; genome-wide; in vitro activity; in vivo; inorganic phosphate; insight; member; novel diagnostics; novel therapeutics; novel vaccines; pathogen; prevent; promoter; protein function; respiratory pathogen; response; therapeutic target; transmission process

Summary/Abstract: Pertussis (aka whooping cough) is a serious reemerging public health problem with incidence estimated at 20 million cases annually and deaths (mostly in infants) at ~200,000 annually. Recent rises in pertussis in countries with high vaccine coverage, such as the United States, correlate with a switch from whole cell (wP) to acellular (aP) pertussis vaccines and are attributed to a larger reservoir of infected individuals composed of adolescents and young adults who were vaccinated (rather than infected) as children. It is now apparent that immunity induced by aP vaccination is not as durable as vaccination by wP vaccination, which is not as durable as immunity induced by infection with Bordetella pertussis, the primary causal agent of pertussis. Moreover, while immunization with wP and aP vaccines provides protection against disease (at least initially), it does not protect against colonization. New vaccines that provide sterilizing, long-lasting immunity are needed. We have discovered a previously uncharacterized signal transduction system (PlrSR) that is required for B. pertussis and the closely-related broad host range pathogen Bordetella bronchiseptica to colonize and persist in the lower respiratory tract (LRT). Our preliminary data support a model in which PlrSR functions as a kinase in response to increased CO2 and low oxygen conditions (reflective of the LRT), resulting in high levels of PlrR-phosphate (PlrR~P) that activate expression of genes including those encoding high-affinity cytochrome oxidases. Our model states that under aerobic conditions, PlrS functions primarily as a phosphatase, and that low levels of PlrR~P are essential for cell viability. We will use genetic, molecular biological, and genome-wide approaches to identify PlrSR-regulated genes, especially those induced only in the LRT, and will determine the roles of PlrSR-dependent gene regulation and of the factors encoded by the regulated genes in virulence. Using genetic approaches, we will determine the role of the PlrS PDC and PAS domains, as well as the predicted kinase and phosphatase activities of PlrS, in the ability of the bacteria to grow in vitro and in the LRT. Using biochemical approaches, we will determine if PlrS is a redox- sensitive heme-containing protein that functions as a kinase under low oxygen conditions and a phosphatase in ambient air, and we will identify DNA sequences to which PlrR~P binds. Our results will be significant because previously unknown PlrSR-dependent virulence factors, and the PlrSR system itself, will be excellent candidates for the development of new component vaccines and targets for the development of new therapeutics. Our results will also advance our understanding NtrYX family proteins (of which PlrSR is a member and which control virulence in other pathogens) function and they will provide insight into how respiratory pathogens, in general, grow in the LRT.

总结/摘要： 百日咳（又名百日咳）是一种严重的重新出现的公共卫生问题， 每年约有2000万例病例，每年约有20万例死亡（主要是婴儿）。最近上涨 在疫苗覆盖率高的国家，如美国，百日咳与从 全细胞（wP）到无细胞（aP）百日咳疫苗，并归因于更大的感染库 由接种疫苗（而不是感染）的青少年和年轻成年人组成的个体， 孩子现在明显的是，由aP疫苗接种诱导的免疫不如由 wP疫苗接种不如百日咳杆菌感染诱导的免疫持久， 百日咳的主要病原体。此外，虽然用wP和aP疫苗免疫提供了 它可以防止疾病（至少在最初），但不能防止殖民化。新的疫苗， 提供杀菌作用，需要持久的免疫力。我们发现了一种以前没有特征的 信号转导系统（PlrSR），其是B所需的。百日咳与近缘广宿主 范围病原体支气管败血波氏杆菌定殖并持续存在于下呼吸道（LRT）中。我们 初步数据支持PlrSR作为激酶响应CO2增加的模型， 低氧条件（反映LRT），导致高水平的PlrR-磷酸盐（PlrRP）， 激活包括编码高亲和力细胞色素氧化酶的基因的表达。我们的模型 指出在有氧条件下，PlrS主要作为磷酸酶发挥作用，低水平的 PlrR~P是细胞生存所必需的。我们将使用遗传学、分子生物学和全基因组技术 鉴定PlrSR调控基因的方法，特别是那些仅在LRT中诱导的基因， 确定PlrSR依赖性基因调控的作用和受调控的基因编码的因子， 毒力基因使用遗传学方法，我们将确定PlrS PDC和PAS的作用， 结构域，以及预测的PlrS激酶和磷酸酶活性，在细菌的能力， 在体外和LRT中生长。使用生物化学方法，我们将确定PlrS是否是氧化还原- 一种敏感的含血红素蛋白，在低氧条件下起激酶作用， 磷酸酶，我们将确定DNA序列的PlrR~P结合。我们的结果将 因为以前未知的PlrSR依赖性毒力因子和PlrSR系统 本身，将是开发新组分疫苗的优秀候选者， 开发新的治疗方法。我们的研究结果也将促进我们对NtrYX家族蛋白的理解 (of PlrSR是其中的一员，并且控制其他病原体中的毒力）的功能，并且它们将 了解呼吸道病原体一般如何在LRT中生长。

项目成果

DegP Initiates Regulated Processing of Filamentous Hemagglutinin in Bordetella bronchiseptica.

• DOI: 10.1128/mbio.01465-21
• 发表时间: 2021-06-29
• 期刊: mBio
• 影响因子: 6.4
• 作者: Johnson RM;Nash ZM;Dedloff MR;Shook JC;Cotter PA
• 通讯作者: Cotter PA