Understanding DNA-binding by type IV pilins: key event during transformation in naturally competent bacteria

了解 IV 型菌毛蛋白的 DNA 结合：自然感受态细菌转化过程中的关键事件

• 批准号: MR/P022197/1
• 负责人: Vladimir Pelicic
• 金额: $ 48.57万
• 依托单位: Imperial College London
• 依托单位国家: 英国
• 项目类别: Research Grant
• 财政年份: 2017
• 资助国家: 英国
• 起止时间: 2017 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=MR%2FP022197%2F1
• 关键词: Understanding; DNA; binding; type; IV

Natural transformation is the biological process during which bacteria capture free DNA from their surroundings and incorporate it into their genomes to become genetically altered by acquiring new genes. This process has important implications for human health because it promotes horizontal transfer of genes between bacteria, which can have devastating medical consequences when it leads to the emergence of bacterial pathogens with increased virulence and/or resistance to antibiotics.The first step during natural transformation, DNA uptake, is mediated by surface filaments named Tff composed of subunits called type IV pilins. These filaments are extremely important because they (i) are almost universal in prokaryotes, and (ii) have been associated with a fascinating array of very different properties, many of which are key for the virulence of bacteria causing important human diseases such as cholera, meningitis, gonorrhoea, diarrhea etc. Studying Tff biology is of great medical interest in this era of rising antibiotic resistance since this could ultimately lead to the development of new antivirulence drugs interfering with Tff assembly or Tff-mediated functions.Until recently, the earliest step in transformation, how Tff interact with DNA was poorly understood because no Tff-associated DNA receptor had ever been identified. Our recent work provided a breakthrough by showing that type IV pilins related to meningococcal ComP display intrinsic DNA-binding ability and bind DNA in a novel fashion. This paves the way to an atomic level understanding of how type IV pilins bind DNA during natural transformation, which is the overarching objective of this research proposal.Using a combination of genetic, biochemical, and structural approaches for which we have extensive experience, we will address the following three aims.Aim 1: determine at an atomic level how ComP pilins bind DNA and how different orthologs are able to distinguish between closely related DNA motifs.Aim 2: characterise the recently identified Pasteurellaceae DNA-binding pilin, unrelated to ComP, to determine how it binds DNA.Aim 3: identify the DNA-binding pilin in competence (pseudo)pili, the most widespread Tff involved in natural transformation.

自然转化是一种生物学过程，在此过程中，细菌从周围环境中捕获游离DNA，并将其整合到基因组中，通过获得新基因进行遗传改变。这一过程对人类健康具有重要影响，因为它促进细菌之间基因的水平转移，当它导致毒力增强和/或对抗生素耐药性增强的细菌病原体出现时，可能会产生毁灭性的医疗后果。自然转化的第一步，DNA吸收，是由名为Tff的表面细丝介导的，由称为IV型菌毛蛋白的亚基组成。这些丝状体非常重要，因为它们（i）在原核生物中几乎是普遍存在的，（ii）与一系列非常不同的特性有关，其中许多特性是细菌毒力的关键，这些细菌会引起重要的人类疾病，如霍乱，脑膜炎，淋病，在这个抗生素耐药性不断上升的时代，研究Tff生物学具有极大的医学意义，因为这可能最终导致新的抗病毒药物干扰Tff组装或Tff介导的功能。直到最近，转化的最早步骤，Tff如何与DNA相互作用还知之甚少，因为还没有鉴定出Tff相关的DNA受体。我们最近的工作提供了一个突破，显示IV型菌毛蛋白相关的脑膜炎球菌ComP显示内在的DNA结合能力，并结合DNA在一个新的方式。这为理解IV型菌毛蛋白在自然转化过程中如何结合DNA铺平了道路，这是本研究提案的首要目标。我们将结合遗传学，生物化学和结构学方法，并在此方面拥有丰富的经验，我们将实现以下三个目标。目标1：在原子水平上确定ComP菌毛蛋白如何结合DNA以及不同的直系同源物如何能够区分密切相关的DNA基序。目的2：目的3：鉴定感受态（假）皮利中的DNA结合菌毛蛋白，该菌毛蛋白是天然转化过程中分布最广的Tff。

项目成果

PilB from Streptococcus sanguinis is a bimodular type IV pilin with a direct role in adhesion.

来自Sanguinis链球菌的PILB是一种双性异的IV型PILIN，在粘附中具有直接作用。

• DOI: 10.1073/pnas.2102092118
• 发表时间: 2021-06-01
• 期刊: Proceedings of the National Academy of Sciences of the United States of America
• 影响因子: 11.1
• 作者: Raynaud C;Sheppard D;Berry JL;Gurung I;Pelicic V
• 通讯作者: Pelicic V

Monoderm bacteria: the new frontier for type IV pilus biology.

Monoderm 细菌：IV 型菌毛生物学的新领域。

• DOI: 10.1111/mmi.14397
• 发表时间: 2019
• 期刊: Molecular microbiology
• 影响因子: 3.6
• 作者: Pelicic V

Full structure/function analysis of all the pilin subunits in a type 4 pilus: a complex of minor pilins in Streptococcus sanguinis mediates binding to glycans

4 型菌毛中所有菌毛蛋白亚基的完整结构/功能分析：血链球菌中的次要菌毛蛋白复合物介导与聚糖的结合

• DOI: 10.1101/2022.08.25.505150
• 发表时间: 2022
• 作者: Shahin M

Structure of a heteropolymeric type 4 pilus from a monoderm bacterium

来自单皮细菌的杂聚型 4 菌毛的结构

• DOI: 10.1101/2023.06.15.545089
• 发表时间: 2023
• 作者: Anger R

Characterization of a glycan-binding complex of minor pilins completes the analysis of Streptococcus sanguinis type 4 pili subunits.

• DOI: 10.1073/pnas.2216237120
• 发表时间: 2023-01-17
• 期刊: PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA
• 影响因子: 11.1
• 作者: Shahin, Meriam;Sheppard, Devon;Raynaud, Claire;Berry, Jamie-Lee;Gurung, Ishwori;Silva, Lisete M.;Feizi, Ten;Liu, Yan;Pelicic, Vladimir
• 通讯作者: Pelicic, Vladimir