Dynamics of the bacterial type IX secretion system and its effect on subgingival biofilm formation by bacteria of the human oral microbiome

细菌 IX 型分泌系统的动态及其对人类口腔微生物组细菌龈下生物膜形成的影响

• 批准号: 10380154
• 负责人: Abhishek Shrivastava
• 金额: $ 24.05万
• 依托单位: ARIZONA STATE UNIVERSITY-TEMPE CAMPUS
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-05-01 至 2023-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10380154
• 关键词: Agar; Bacteria; Bacterial Adhesins; Bacterial Genes; Behavior; Biological Assay; Capnocytophaga; Cell Adhesion; Cell Aggregation; Cell secretion; Cell surface; Cells; Chemicals; Communities; Complex; Coupled; Cues; Data; Development; Elasticity; Environment; Enzymes; Escherichia coli; Exhibits; Forsythia; Frequencies; Gene Expression; Gene Expression Regulation; Genes; Genetic; Genetic Screening; Genotype; Human; Hydrogels; Imaging Device; Immune response; Lead; Lys-gingipain; Methods; Microbe; Microbial Biofilms; Modulus; Monitor; Motor; Movement; Mucins; Mucous body substance; Mutation; Myxococcus xanthus; Oral; Output; Pathway interactions; Peptide Hydrolases; Periodontal Diseases; Phase; Pilum; Plasma Proteins; Population; Porphyromonas gingivalis; Process; Protein Secretion; Proteins; Proteomics; Pseudomonas aeruginosa; Publishing; Pump; Regulatory Pathway; Reporting; Research; Risk; Role; Rotation; Salivary; Salmonella enterica; Sensory; Serum; Signal Pathway; Signaling Protein; Speed; Suction; Surface; System; Testing; Tissues; Type III Secretion System Pathway; Virulence Factors; biophysical techniques; cell motility; gingipain; gliding bacteria; human microbiota; human pathogen; hydrodynamic flow; mechanical force; mechanical stimulus; mechanotransduction; microbiome components; mutant; novel; oral bacteria; oral biofilm; oral microbiome; pressure; prevent; response; sensor; subgingival biofilm; tool; transcriptome sequencing; transposon sequencing

Project Summary/Abstract The bacterial Type IX protein secretion system (T9SS), which is a recently discovered protein secretion system, is found in bacteria that are a part of human subgingival biofilms. So far, it has been shown that T9SS is functional in Porphyromonas gingivalis [1, 2], Tannerella forsythia [3] and Capnocytophaga sp. [4]. P. gingivalis Arg-gingipain (Rgp) and Lys-gingipain (Kgp) are secreted via T9SS. Rgp and Kgp increase the risk of periodontal diseases by disrupting the host immune response and degradation of the host tissue and plasma proteins. T9SS is associated with a rotary motor and its dynamics enables secretion of cell-surface adhesins, known virulence factors and enzymes. It is also required for movement of bacteria over a surface by a process called gliding motility. Cell adhesion, surface navigation and motility are important for the formation of many bacterial biofilms [5]. Recent reports suggest that bacterial motility machineries enable sensing of external surfaces via a process called as mechanosensing [6, 7]. Human pathogens such as Escherichia coli and Salmonella enterica sense external surfaces via the flagellar motor-Type III secretion system machinery [6], while Pseudomonas aeruginosa and Myxococcus xanthus sense external surface via the Type IV pili machinery [7, 8]. Our preliminary and published results suggest that gliding bacteria with T9SS have the ability to sense external surfaces and viscous environments [9]. Such bacteria do not have the flagellar motor or the type IV pili. Amongst bacteria of the human oral microbiome, members of Capnocytophaga genus, which are present in human subgingival biofilms [10], are an attractive system for answering important questions related to biofilm formation, surface-sensing, and hydrodynamics. These bacteria have the T9SS and they exhibit gliding motility over external surfaces. Recently, genetic tools were developed for their manipulation [4]. I propose to study Capnocytophaga gingivalis, which is an important, yet understudied bacterium present in human subgingival biofilms [4, 10]. We discovered that the T9SS of C. gingivalis is powered by a novel rotary motor. Also, we found that C. gingivalis cells use adhesins secreted by T9SS to aggregate and form groups. Such aggregates exhibit gliding motility over external surfaces. Our data suggests that gliding is powered by the same rotary motor that powers secretion via T9SS. Gliding motility, which is the output of the dynamics of T9SS, will be used to assay for gene regulation and for development of genetic screens. Such screens will be targeted towards identification of proteins involved in sensory transduction by oral bacteria. Overall, dynamics of T9SS and role of proteins secreted by T9SS will be studied in the context of subgingival biofilm formation.

项目摘要/摘要 细菌型IX蛋白分泌系统（T9SS），这是最近发现的蛋白质分泌 在细菌中发现的系统是人类副生物膜的一部分。到目前为止，已经表明 T9SS在牙龈卟啉单胞菌[1，2]，Tannerella Forsythia [3]和capnocytophaga sp中起作用。 [4]。牙龈疟原虫Arg-gyipain（RGP）和Lys-ingipain（KGP）通过T9SS分泌。 RGP和KGP 通过破坏宿主免疫反应和降解来增加牙周疾病的风险 宿主组织和血浆蛋白。 T9SS与旋转电动机有关，其动力学启用 细胞表面粘附，已知毒力因子和酶的分泌。也需要 细菌通过称为滑行运动的过程在表面上运动。细胞粘附，表面 导航和运动对于形成许多细菌生物膜非常重要[5]。最近的报告 建议细菌运动机制可以通过称为AS的过程来感知外表面 机械感应[6，7]。大肠杆菌和沙门氏菌等人类病原体 通过鞭毛运动型III分泌系统机械的外表面[6]，而假单胞菌 铜绿和粘膜Xanthus Xanthus通过IV型Pili机械感知外表面[7，8]。我们的 初步和发表的结果表明，使用T9S的滑行细菌具有感知的能力 外表面和粘性环境[9]。这样的细菌没有鞭毛电机或类型 iv pili。在人口腔微生物组的细菌中 存在于人类副生物膜[10]中，是回答重要问题的有吸引力的系统 与生物膜形成，表面感应和流体动力学有关。这些细菌具有T9S和 它们在外表面上表现出滑动运动。最近，为他们开发了遗传工具 操纵[4]。我建议研究囊孢子的牙龈，这是一个重要但又研究的 人类副生物膜中存在的细菌[4，10]。我们发现牙龈梭菌的T9S是 由新型旋转电动机提供动力。另外，我们发现牙龈梭菌细胞使用T9SS分泌的粘附素 汇总和形成组。这样的聚集体在外表面上表现出滑行运动。我们的数据 表明滑行是由通过T9SS供电的相同旋转电动机提供动力的。滑翔 运动性是T9SS动力学的输出，将用于测定基因调节和 遗传筛查的发展。此类筛选将针对鉴定涉及的蛋白质 在口腔细菌的感觉转导。总体而言，T9SS的动态和蛋白质的作用 T9SS将在尺寸生物膜形成的背景下进行研究。