PlrSR-dependent Signal Transduction in Bordetella Virulence

博德特氏菌毒力中 PlrSR 依赖性信号转导

基本信息

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
项目类别：
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

项目摘要

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.

摘要/摘要：百日咳（又名百日咳）是一个严重的公共卫生问题，估计发病率每年有2000万例，每年约20万。最近上升疫苗覆盖范围高的国家（例如美国）的百日咳与从全细胞（WP）到细胞（AP）百日咳疫苗，归因于较大的感染库由青少年和年轻人组成的个人，他们被接种（而不是感染）为孩子们。现在显然，AP疫苗接种引起的免疫力不如疫苗接种 WP疫苗接种，不如bordetella budtussis感染的免疫力耐用，百日咳的主要因果因素。此外，虽然WP和AP疫苗的免疫提供保护疾病（至少在最初），它不能防止殖民化。新疫苗需要进行消毒，持久的免疫力。我们发现了一个以前没有特色的百日咳和密切相关的宿主所需的信号转导系统（PLRSR）范围病原体支气管肽范围内定植并持续在下呼吸道（LRT）中。我们的初步数据支持一个模型，其中PLRSR作为激酶，以响应增加的CO2和低氧条件（反射LRT），导致高水平的PLRR-磷酸（PLRR〜P）激活基因的表达，包括编码高亲和力细胞色素氧化酶的基因。我们的模型指出，在有氧条件下，PLRS主要起作用为磷酸酶，低水平 PLRR〜P对于细胞活力至关重要。我们将使用遗传，分子生物学和全基因组鉴定PLRSR调节的基因，尤其是仅在LRT中诱导的基因的方法，并将确定PLRSR依赖性基因调节的作用以及由调节的因素的作用毒力中的基因。使用遗传方法，我们将确定PLRS PDC和PAS的作用 PLR的域以及预测的激酶和磷酸酶活性在细菌的能力方面在体外和LRT中生长。使用生化方法，我们将确定PLR是否是氧化还原敏感的含血红素的蛋白质，在低氧条件下充当激酶，A 环境空气中的磷酸酶，我们将确定plrr〜p结合的DNA序列。我们的结果将会要重要，因为以前未知的PLRSR依赖性毒力因子和PLRSR系统本身，将是开发新组件疫苗和目标的出色候选人开发新治疗学。我们的结果还将提高我们的了解Ntryx家庭蛋白（其中PLRSR是成员，并且控制其他病原体中的毒力）功能，它们将提供有关通常在LRT中如何生长的呼吸道病原体的见解。