Role of the Listeria protein InlC and its host ligand Tuba in cell-cell spread

李斯特菌蛋白 InlC 及其宿主配体 Tuba 在细胞间传播中的作用

基本信息

  • 批准号:
    8040584
  • 负责人:
  • 金额:
    $ 9.99万
  • 依托单位:
  • 依托单位国家:
    美国
  • 项目类别:
  • 财政年份:
    2008
  • 资助国家:
    美国
  • 起止时间:
    2008-06-19 至 2011-05-31
  • 项目状态:
    已结题

项目摘要

DESCRIPTION (provided by applicant): Listeria monocytogenes is a food-borne, intracellular pathogen capable of causing gastroenteritis, meningitis, and abortions. Critical for Listeria virulence is its ability to replicate within mammalian cells, and spread from one cell to another through an actin-dependent motility process that allows bacteria to remain within the protective environment of the host cytosol. Cell-cell spread is initiated by the bacterial surface protein ActA, which stimulates the formation of F-actin 'comet tails' that propel Listeria through the cytoplasm. Rocketing bacteria contact the host cell plasma membrane and induce the formation of Listeria-containing protrusions that are engulfed by neighboring cells. Although ActA-mediated actin polymerization is well understood, it remains unclear whether additional bacterial factors promote protrusion formation and intercellular spread by acting after the step of comet tail formation. Our preliminary data indicate that the secreted Listeria virulence protein InlC is needed for efficient cell-cell spread, but is not essential for bacterial-induced F-actin assembly. Importantly, InlC promotes activation (phosphorylation) of host ERM proteins, a family of cytoskeletal regulatory proteins known to participate in Listeria-induced protrusion formation and spread. Moreover, InlC localizes to a subset of comet tails that also contain the ERM protein ezrin. Through a yeast two-hybrid screen, the human adaptor protein Tuba was identified as a ligand of InlC. Like InlC, Tuba co-localizes with ezrin in a subset of comet tails. Tuba contains several functional domains, including SH3 domains that engage ezrin (SH35) or InlC (SH36). Importantly, RNA interference studies indicate that Tuba antagonizes cell-cell spread of Listeria. InlC promotes spreading by relieving the inhibitory effect of Tuba, possibly by impairing binding of human SH36 ligands to Tuba. Based on these results, it seems likely that InlC, Tuba, and ezrin act together to promote cell-cell spread. Tuba might influence spreading by affecting ERM proteins or other effectors. The long-term goal of this project is to understand the molecular mechanism by which InlC promotes intercellular dissemination of Listeria. The Specific Aims in this proposal are directed towards understanding how InlC, ERM proteins, Tuba, and Tuba ligands control cell-cell spread. 1. Determine if InlC and/or Tuba affect formation of F-actin tails or protrusions. 2. Determine if InlC and/or Tuba control spreading through ERM protein phosphorylation. 3. Evaluate the role of human SH36 ligands in intercellular spread of Listeria. The above objectives will be addressed through a variety of approaches, including immunofluorescence microscopy, evaluation of ERM protein phoshorylation through Western blotting, and RNA interference to investigate the function of Tuba and Tuba ligands in cell-cell spread. Listeria monocytogenes is a food-borne, intracellular bacterium that causes serious illnesses leading to meningitis or abortion. Listeria evades the immune system by spreading from one human cell to another, while remaining within the protective environment of the host cytosol. The goal of this research is to identify bacterial and human components that mediate Listeria spreading, and which may be suitable as drug targets.
描述(由申请人提供):单核细胞增生李斯特氏菌是一种食源性细胞内病原体,能够引起胃肠炎、脑膜炎和流产。李斯特菌毒力的关键在于其在哺乳动物细胞内复制的能力,并通过肌动蛋白依赖性运动过程从一个细胞传播到另一个细胞,该过程使细菌能够保留在宿主细胞质的保护环境中。细胞间的传播是由细菌表面蛋白 ActA 启动的,它刺激 F-肌动蛋白“彗尾”的形成,推动李斯特菌穿过细胞质。火箭细菌接触宿主细胞质膜并诱导形成含有李斯特菌的突出物,这些突出物被邻近的细胞吞噬。尽管 ActA 介导的肌动蛋白聚合已被充分了解,但仍不清楚其他细菌因子是否通过在彗尾形成步骤后起作用来促进突起形成和细胞间扩散。我们的初步数据表明,分泌的李斯特菌毒力蛋白InlC对于有效的细胞间传播是必需的,但对于细菌诱导的F-肌动蛋白组装来说并不是必需的。重要的是,InlC 促进宿主 ERM 蛋白的激活(磷酸化),这是一个已知参与李斯特菌诱导的突起形成和扩散的细胞骨架调节蛋白家族。此外,InlC 定位于也含有 ERM 蛋白埃兹蛋白的彗尾子集。通过酵母双杂交筛选,人衔接蛋白Tuba被鉴定为InlC的配体。与 InlC 一样,Tuba 与埃兹蛋白共定位于彗尾的一个子集中。 Tuba 包含多个功能域,包括与 ezrin (SH35) 或 InlC (SH36) 结合的 SH3 域。重要的是,RNA 干扰研究表明 Tuba 可以拮抗李斯特菌的细胞间传播。 InlC 通过减轻 Tuba 的抑制作用(可能是通过削弱人 SH36 配体与 Tuba 的结合)来促进扩散。基于这些结果,InlC、Tuba 和 ezrin 很可能共同作用,促进细胞间的扩散。 Tuba 可能通过影响 ERM 蛋白或其他效应物来影响传播。 该项目的长期目标是了解InlC促进李斯特菌细胞间传播的分子机制。该提案的具体目标是了解 InlC、ERM 蛋白、Tuba 和 Tuba 配体如何控制细胞间扩散。 1. 确定 InlC 和/或 Tuba 是否影响 F-肌动蛋白尾部或突起的形成。 2. 确定 InlC 和/或 Tuba 是否通过 ERM 蛋白磷酸化控制扩散。 3. 评估人SH36配体在李斯特菌细胞间传播中的作用。 上述目标将通过多种方法来实现,包括免疫荧光显微镜、通过蛋白质印迹评估 ERM 蛋白磷酸化,以及通过 RNA 干扰研究 Tuba 和 Tuba 配体在细胞间扩散中的功能。 单核细胞增生李斯特菌是一种食源性细胞内细菌,可引起严重疾病,导致脑膜炎或流产。李斯特菌通过从一个人体细胞传播到另一个人体细胞来逃避免疫系统,同时保留在宿主细胞质的保护环境中。这项研究的目标是确定介导李斯特菌传播的细菌和人体成分,并可能适合作为药物靶点。

项目成果

期刊论文数量(1)
专著数量(0)
科研奖励数量(0)
会议论文数量(0)
专利数量(0)
The bacterial virulence factor InlC perturbs apical cell junctions and promotes cell-to-cell spread of Listeria.
  • DOI:
    10.1038/ncb1964
  • 发表时间:
    2009-10
  • 期刊:
  • 影响因子:
    21.3
  • 作者:
    Rajabian, Tina;Gavicherla, Balramakrishna;Heisig, Martin;Mueller-Altrock, Stefanie;Goebel, Werner;Gray-Owen, Scott D.;Ireton, Keith
  • 通讯作者:
    Ireton, Keith
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KEITH Patrick IRETON其他文献

KEITH Patrick IRETON的其他文献

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{{ truncateString('KEITH Patrick IRETON', 18)}}的其他基金

Molecular Mechanism of cell-cell spread of Listeria in polarized epithelial cells
李斯特菌在极化上皮细胞中细胞间传播的分子机制
  • 批准号:
    7985826
  • 财政年份:
    2010
  • 资助金额:
    $ 9.99万
  • 项目类别:
Molecular Mechanism of cell-cell spread of Listeria in polarized epithelial cells
李斯特菌在极化上皮细胞中细胞间传播的分子机制
  • 批准号:
    8513124
  • 财政年份:
    2010
  • 资助金额:
    $ 9.99万
  • 项目类别:
Molecular Mechanism of cell-cell spread of Listeria in polarized epithelial cells
李斯特菌在极化上皮细胞中细胞间传播的分子机制
  • 批准号:
    8307716
  • 财政年份:
    2010
  • 资助金额:
    $ 9.99万
  • 项目类别:
Molecular Mechanism of cell-cell spread of Listeria in polarized epithelial cells
李斯特菌在极化上皮细胞中细胞间传播的分子机制
  • 批准号:
    8122184
  • 财政年份:
    2010
  • 资助金额:
    $ 9.99万
  • 项目类别:
Role of the Listeria protein InlC and its host ligand Tuba in cell-cell spread
李斯特菌蛋白 InlC 及其宿主配体 Tuba 在细胞间传播中的作用
  • 批准号:
    7362720
  • 财政年份:
    2008
  • 资助金额:
    $ 9.99万
  • 项目类别:
Role of the Listeria protein InlC and its host ligand Tuba in cell-cell spread
李斯特菌蛋白 InlC 及其宿主配体 Tuba 在细胞间传播中的作用
  • 批准号:
    7640660
  • 财政年份:
    2008
  • 资助金额:
    $ 9.99万
  • 项目类别:
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