Structure and Function of Pathogenesis-Associated Bacterial Structures by Electron Cryotomography

通过电子冷冻断层扫描研究发病机制相关细菌结构的结构和功能

基本信息

  • 批准号:
    10604243
  • 负责人:
  • 金额:
    $ 36.95万
  • 依托单位:
  • 依托单位国家:
    美国
  • 项目类别:
  • 财政年份:
    2016
  • 资助国家:
    美国
  • 起止时间:
    2016-09-23 至 2027-03-31
  • 项目状态:
    未结题

项目摘要

Project Summary Pathogenic bacteria employ specialized secretion systems to identify and interact with host cells and to exchange genetic information through horizontal gene transfer. These machines are attractive drug targets because they are surface-exposed, widely conserved, and specific for pathogenicity. Unfortunately, however, the structures of many of these critical systems remain poorly understood. Here we describe how we will continue to use electron cryotomography (cryoET) to dissect the structures and functions of pathogenic nanomachines. CryoET is a revolutionary imaging technique with the power to reveal native structures inside intact cells in 3D with macromolecular (2-5 nm) resolution. Subtomogram averaging of identical structures from one or more cryotomograms can push this resolution to better than 1 nm in the most favorable cases, enabling components to be placed in their context in the complete machine. My group has pioneered the development of this revolutionary imaging technology, and in just under four years of our first award period, we have used cryoET to produce tens of new structures of pathogenic secretion systems and build architectural models of key systems belonging to the type IV pilus (T4P), type VI secretion system (T6SS) and type IV secretion system (T4SS) families, producing a flood of new mechanistic insights. By exploiting new cryoET technologies we have just developed in the past couple years, here we propose to extend our work in the next award period to different functional states of these complexes, key related systems, and a new target: the pathogenic type IX secretion system (T9SS). In addition, we will push the whole body of work to higher resolution. For each target, we will image the entire, intact structure in situ. In most cases, this will be the first high-resolution imaging of these structures. We will then combine subtomogram averaging with difference analysis of mutants in which individual components are knocked out or tagged with additional density in order to produce architectural models of the complexes. In cases where atomic models of components (or homologs) are available, we will dock them into our maps to produce pseudo-atomic models of each machine. By comparing these structures with those of non-pathogenic relatives (solved previously or in the proposed work), we aim to identify adaptations underlying virulence functions. We will also apply state-of-the-art cryogenic correlated light and electron microscopy (cryo-CLEM) to guide cryogenic focused ion beam (FIB) milling to enable us to image pathogenic secretion systems in action: in bacterial cells infecting eukaryotic hosts. This will provide the first such images of critical human pathogens, which we expect to provide invaluable insights into the operation of their virulence machinery in vivo. Together, we expect this project to produce a detailed mechanistic picture of the T4SS, T4P, and T9SS nanomachines that mediate pathogenesis, an important first step in identifying therapeutic targets in the future.
项目摘要 病原菌利用专门的分泌系统来识别宿主细胞并与宿主细胞相互作用, 通过水平基因转移交换遗传信息。这些机器是有吸引力的药物靶点 因为它们是表面暴露的,广泛保守的,并且对致病性具有特异性。然而不幸的是, 对其中许多关键系统的结构仍然知之甚少。在这里,我们描述我们将如何 继续使用电子冷冻断层扫描(cryoET)来解剖致病性组织的结构和功能, 纳米机器CryoET是一种革命性的成像技术,具有揭示内部天然结构的能力 完整的细胞在3D与大分子(2-5 nm)的分辨率。相同结构的子断层图像平均值, 在最有利的情况下,一个或多个冷冻断层照片可以将该分辨率推到优于1 nm, 这些组件将被放置在整个机器中的上下文中。我的团队率先开发了 这项革命性的成像技术,在我们第一个奖项期的不到四年时间里,我们已经使用了 cryoET产生了数十种致病分泌系统的新结构,并建立了 属于IV型菌毛(T4 P)、VI型分泌系统(T6 SS)和IV型分泌的关键系统 系统(T4 SS)家族,产生了大量新的机械见解。通过开发新的冷冻ET技术 我们在过去的几年里刚刚发展起来,在此我们建议在下一个奖励期内继续我们的工作 这些复合物的不同功能状态,关键相关系统和一个新的目标:致病性IX型 分泌系统(T9 SS)。此外,我们将推动整个工作的更高分辨率。对于每个目标, 我们将在原位对整个完整的结构进行成像。在大多数情况下,这将是第一个高分辨率成像的 这些结构。然后,我们将联合收割机子断层图像平均与突变体的差异分析相结合, 单个组件被敲除或标记有额外的密度,以产生建筑 复合体的模型。在组件(或同系物)的原子模型可用的情况下,我们将 将它们对接到我们的地图中,以生成每台机器的伪原子模型。通过比较这些结构 与那些非致病性的亲属(以前解决或在拟议的工作),我们的目标是确定 适应潜在的毒力功能。我们还将应用最先进的低温相关光, 电子显微镜(cryo-CLEM)引导低温聚焦离子束(FIB)铣削,使我们能够成像 致病分泌系统的作用:在细菌细胞感染真核宿主。这将提供第一个 这些关键的人类病原体的图像,我们希望提供宝贵的见解的运作, 它们在体内的毒力机制。总之,我们希望这个项目能产生一个详细的机械图, 介导发病机制的T4 SS、T4 P和T9 SS纳米机器,这是识别 未来的治疗目标。

项目成果

期刊论文数量(28)
专著数量(0)
科研奖励数量(0)
会议论文数量(0)
专利数量(0)
Programmed Flagellar Ejection in Caulobacter crescentus Leaves PL-subcomplexes.
  • DOI:
    10.1016/j.jmb.2021.167004
  • 发表时间:
    2021-06-25
  • 期刊:
  • 影响因子:
    5.6
  • 作者:
    Kaplan, Mohammed;Wang, Yuhang;Chreifi, Georges;Zhang, Lujia;Chang, Yi-Wei;Jensen, Grant J.
  • 通讯作者:
    Jensen, Grant J.
Structure of Anabaena flos-aquae gas vesicles revealed by cryo-ET.
  • DOI:
    10.1016/j.str.2023.03.011
  • 发表时间:
    2023-05-04
  • 期刊:
  • 影响因子:
    5.7
  • 作者:
    Dutka, Przemysaw;Metskas, Lauren Ann;Hurt, Robert C.;Salahshoor, Hossein;Wang, Ting-Yu;Malounda, Dina;Lu, George J.;Chou, Tsui-Fen;Shapiro, Mikhail G.;Jensen, Grant J.
  • 通讯作者:
    Jensen, Grant J.
The Atlas of Bacterial & Archaeal Cell Structure: an Interactive Open-Access Microbiology Textbook.
Subtomogram averaging for biophysical analysis and supramolecular context.
  • DOI:
    10.1016/j.yjsbx.2022.100076
  • 发表时间:
    2022
  • 期刊:
  • 影响因子:
    2.9
  • 作者:
    Metskas, Lauren Ann;Wilfong, Rosalie;Jensen, Grant J.
  • 通讯作者:
    Jensen, Grant J.
Loss of the Bacterial Flagellar Motor Switch Complex upon Cell Lysis.
  • DOI:
    10.1128/mbio.00298-21
  • 发表时间:
    2021-06-29
  • 期刊:
  • 影响因子:
    6.4
  • 作者:
    Kaplan M;Tocheva EI;Briegel A;Dobro MJ;Chang YW;Subramanian P;McDowall AW;Beeby M;Jensen GJ
  • 通讯作者:
    Jensen GJ
{{ item.title }}
{{ item.translation_title }}
  • DOI:
    {{ item.doi }}
  • 发表时间:
    {{ item.publish_year }}
  • 期刊:
  • 影响因子:
    {{ item.factor }}
  • 作者:
    {{ item.authors }}
  • 通讯作者:
    {{ item.author }}

数据更新时间:{{ journalArticles.updateTime }}

{{ item.title }}
  • 作者:
    {{ item.author }}

数据更新时间:{{ monograph.updateTime }}

{{ item.title }}
  • 作者:
    {{ item.author }}

数据更新时间:{{ sciAawards.updateTime }}

{{ item.title }}
  • 作者:
    {{ item.author }}

数据更新时间:{{ conferencePapers.updateTime }}

{{ item.title }}
  • 作者:
    {{ item.author }}

数据更新时间:{{ patent.updateTime }}

GRANT J JENSEN其他文献

GRANT J JENSEN的其他文献

{{ item.title }}
{{ item.translation_title }}
  • DOI:
    {{ item.doi }}
  • 发表时间:
    {{ item.publish_year }}
  • 期刊:
  • 影响因子:
    {{ item.factor }}
  • 作者:
    {{ item.authors }}
  • 通讯作者:
    {{ item.author }}

{{ truncateString('GRANT J JENSEN', 18)}}的其他基金

Expansion of the ‘Getting Started in Cryo-EM’ course into a comprehensive theory and practice curriculum
将“Cryo-EM 入门”课程扩展为综合理论和实践课程
  • 批准号:
    10223807
  • 财政年份:
    2021
  • 资助金额:
    $ 36.95万
  • 项目类别:
Expansion of the 'Getting Started in Cryo-EM' course into a comprehensive theory and practice curriculum
将“冷冻电镜入门”课程扩展为综合理论和实践课程
  • 批准号:
    10834296
  • 财政年份:
    2021
  • 资助金额:
    $ 36.95万
  • 项目类别:
Expansion of the 'Getting Started in Cryo-EM' course into a comprehensive theory and practice curriculum
将“冷冻电镜入门”课程扩展为综合理论和实践课程
  • 批准号:
    10798674
  • 财政年份:
    2021
  • 资助金额:
    $ 36.95万
  • 项目类别:
Expansion of the ‘Getting Started in Cryo-EM’ course into a comprehensive theory and practice curriculum
将“Cryo-EM 入门”课程扩展为综合理论和实践课程
  • 批准号:
    10437759
  • 财政年份:
    2021
  • 资助金额:
    $ 36.95万
  • 项目类别:
Imaging large macromolecular complexes inside cells with electron cryotomography
使用电子冷冻断层扫描对细胞内的大分子复合物进行成像
  • 批准号:
    10013429
  • 财政年份:
    2017
  • 资助金额:
    $ 36.95万
  • 项目类别:
Structure and function of pathogenesis-associated bacterial structures by electron cryotomography
通过电子冷冻断层扫描研究发病机制相关细菌结构的结构和功能
  • 批准号:
    9765150
  • 财政年份:
    2016
  • 资助金额:
    $ 36.95万
  • 项目类别:
Structure and function of pathogenesis-associated bacterial structures by electron cryotomography
通过电子冷冻断层扫描研究发病机制相关细菌结构的结构和功能
  • 批准号:
    9357518
  • 财政年份:
    2016
  • 资助金额:
    $ 36.95万
  • 项目类别:
Structural Basis of Chemoreception
化学感受的结构基础
  • 批准号:
    8643266
  • 财政年份:
    2013
  • 资助金额:
    $ 36.95万
  • 项目类别:
Structural Basis of Chemoreception
化学感受的结构基础
  • 批准号:
    9022486
  • 财政年份:
    2013
  • 资助金额:
    $ 36.95万
  • 项目类别:
Structural Basis of Chemoreception
化学感受的结构基础
  • 批准号:
    8507076
  • 财政年份:
    2013
  • 资助金额:
    $ 36.95万
  • 项目类别:

相似海外基金

How tensins transform focal adhesions into fibrillar adhesions and phase separate to form new adhesion signalling hubs.
张力蛋白如何将粘着斑转化为纤维状粘连并相分离以形成新的粘连信号中枢。
  • 批准号:
    BB/Y004841/1
  • 财政年份:
    2024
  • 资助金额:
    $ 36.95万
  • 项目类别:
    Research Grant
Defining a role for non-canonical mTORC1 activity at focal adhesions
定义非典型 mTORC1 活性在粘着斑中的作用
  • 批准号:
    BB/Y001427/1
  • 财政年份:
    2024
  • 资助金额:
    $ 36.95万
  • 项目类别:
    Research Grant
How tensins transform focal adhesions into fibrillar adhesions and phase separate to form new adhesion signalling hubs.
张力蛋白如何将粘着斑转化为纤维状粘连并相分离以形成新的粘连信号中枢。
  • 批准号:
    BB/Y005414/1
  • 财政年份:
    2024
  • 资助金额:
    $ 36.95万
  • 项目类别:
    Research Grant
Development of a single-use, ready-to-use, sterile, dual chamber, dual syringe sprayable hydrogel to prevent postsurgical cardiac adhesions.
开发一次性、即用型、无菌、双室、双注射器可喷雾水凝胶,以防止术后心脏粘连。
  • 批准号:
    10669829
  • 财政年份:
    2023
  • 资助金额:
    $ 36.95万
  • 项目类别:
Regulating axon guidance through local translation at adhesions
通过粘连处的局部翻译调节轴突引导
  • 批准号:
    10587090
  • 财政年份:
    2023
  • 资助金额:
    $ 36.95万
  • 项目类别:
Improving Maternal Outcomes of Cesarean Delivery with the Prevention of Postoperative Adhesions
通过预防术后粘连改善剖宫产的产妇结局
  • 批准号:
    10821599
  • 财政年份:
    2023
  • 资助金额:
    $ 36.95万
  • 项目类别:
Regulating axon guidance through local translation at adhesions
通过粘连处的局部翻译调节轴突引导
  • 批准号:
    10841832
  • 财政年份:
    2023
  • 资助金额:
    $ 36.95万
  • 项目类别:
Prevention of Intraabdominal Adhesions via Release of Novel Anti-Inflammatory from Surface Eroding Polymer Solid Barrier
通过从表面侵蚀聚合物固体屏障中释放新型抗炎剂来预防腹内粘连
  • 批准号:
    10532480
  • 财政年份:
    2022
  • 资助金额:
    $ 36.95万
  • 项目类别:
I-Corps: A Sprayable Tissue-Binding Hydrogel to Prevent Postsurgical Cardiac Adhesions
I-Corps:一种可喷雾的组织结合水凝胶,可防止术后心脏粘连
  • 批准号:
    10741261
  • 财政年份:
    2022
  • 资助金额:
    $ 36.95万
  • 项目类别:
Sprayable Polymer Blends for Prevention of Site Specific Surgical Adhesions
用于预防特定部位手术粘连的可喷涂聚合物共混物
  • 批准号:
    10674894
  • 财政年份:
    2022
  • 资助金额:
    $ 36.95万
  • 项目类别:
{{ showInfoDetail.title }}

作者:{{ showInfoDetail.author }}

知道了