Molecular analysis of actin-based motility of Burkholderia spp.

伯克霍尔德氏菌基于肌动蛋白的运动的分子分析。

• 批准号: BB/E021212/1
• 负责人: Mark Stevens
• 金额: $ 51.06万
• 依托单位: The Pirbright Institute
• 依托单位国家: 英国
• 项目类别: Research Grant
• 财政年份: 2007
• 资助国家: 英国
• 起止时间: 2007 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=BB%2FE021212%2F1
• 关键词: Molecular; analysis; actin; based; motility

Burkholderia pseudomallei is a bacterial pathogen that causes melioidosis, a severe and emerging disease of animals and humans that is endemic in the subtropics. In common with some members of the genera Listeria, Shigella, Rickettsia and Mycobacterium, B. pseudomallei has evolved the remarkable ability to enter host cells and propel itself within and between cells by hijacking the cellular machinery that assembles actin, a key structural component of the cell skeleton. In all cases so far examined, the ability of these organisms to fire themselves into adjacent cells on actin 'rockets' is required to cause disease, presumably as it aids cell-to-cell spread of the bacteria while evading host immune surveillance. The molecular basis of the actin-dependent movement of Listeria and Shigella have been extensively studied, partly to understand how the organisms cause disease, but also because it provides insights into how actin assembly is orchestrated and regulated inside cells. Actin is one of the most abundant proteins in our cells and control of actin assembly is essential for a multitude of processes, including cell migration during embryo development, wound healing and invasive cancer, as well as the movement and ability of immune cells to fight infection. With BBSRC funding we recently identified a bacterial factor that is required for actin-based motility of B. pseudomallei (BimA), as well as similar functional proteins in related Burkholderia species. Though we have established that these factors bind actin and promote its assembly in the test tube and inside host cells, the mechanism by which they do this is not understood. In this proposal we seek to: 1. Identify regions of the BimA protein that are required for actin binding and assembly. 2. Determine whether modifications of BimA by addition of phosphate and/or sugar groups occur inside cells, and investigate their functional consequences. 3. Examine the complex assembled by BimA at the bacterial surface and the protein-protein interactions that occur within it. 4. Detect host cell proteins in B. pseudomallei-induced actin tails inside cells and determine their functional relevance. 5. Determine if BimA proteins from different Burkholderia species promote the formation of complexes with differing compositions. Taken together, we believe that these experiments will explain the mode of action of key microbial virulence factors and produce important new knowledge on the mechanisms by which actin dynamics are controlled in our cells.

类鼻疽伯克霍尔德氏菌是一种引起类鼻疽的细菌病原体，类鼻疽是一种严重的新出现的动物和人类疾病，在亚热带地区流行。与李斯特菌属、志贺氏菌属、立克次体属和分枝杆菌属的某些成员相同，B。类鼻疽已进化出进入宿主细胞并通过劫持组装肌动蛋白（细胞骨架的关键结构组分）的细胞机器而在细胞内和细胞间推进自身的非凡能力。在迄今为止研究的所有情况下，这些生物体在肌动蛋白“火箭”上将自己发射到相邻细胞中的能力是导致疾病所必需的，大概是因为它有助于细菌的细胞间传播，同时逃避宿主的免疫监视。李斯特菌和志贺氏菌的肌动蛋白依赖性运动的分子基础已被广泛研究，部分原因是为了了解生物体如何引起疾病，但也因为它提供了对肌动蛋白组装如何在细胞内协调和调节的见解。肌动蛋白是我们细胞中最丰富的蛋白质之一，肌动蛋白组装的控制对于许多过程至关重要，包括胚胎发育期间的细胞迁移，伤口愈合和侵袭性癌症，以及免疫细胞对抗感染的运动和能力。在BBSRC的资助下，我们最近发现了一种细菌因子，它是B基于肌动蛋白的运动所必需的。类鼻疽杆菌（比马），以及相关伯克霍尔德氏菌属物种中的类似功能蛋白。虽然我们已经确定这些因子结合肌动蛋白并促进其在试管和宿主细胞内的组装，但它们这样做的机制尚不清楚。在本提案中，我们寻求：1。确定肌动蛋白结合和组装所需的比马蛋白区域。2.确定通过添加磷酸和/或糖基修饰比马是否发生在细胞内，并研究其功能后果。3.检测比马在细菌表面组装的复合物及其内部发生的蛋白质-蛋白质相互作用。检测B中的宿主细胞蛋白。pseudomallei诱导的细胞内的肌动蛋白尾巴，并确定其功能的相关性。5.确定来自不同伯克霍尔德菌属种的比马蛋白是否促进具有不同组成的复合物的形成。综上所述，我们相信这些实验将解释关键微生物毒力因子的作用模式，并产生关于我们细胞中肌动蛋白动力学控制机制的重要新知识。

项目成果

Autotransporters and Their Role in the Virulence of Burkholderia pseudomallei and Burkholderia mallei.

• DOI: 10.3389/fmicb.2011.00151
• 发表时间: 2011
• 期刊: Frontiers in microbiology
• 影响因子: 5.2
• 作者: Lazar Adler NR;Stevens JM;Stevens MP;Galyov EE
• 通讯作者: Galyov EE

Burkholderia pseudomallei-induced cell fusion in U937 macrophages can be inhibited by monoclonal antibodies against host cell surface molecules

针对宿主细胞表面分子的单克隆抗体可以抑制鼻疽伯克霍尔德氏菌诱导的 U937 巨噬细胞的细胞融合

• DOI: 10.1016/j.micinf.2011.06.007
• 发表时间: 2011
• 期刊: Microbes and Infection
• 影响因子: 5.8
• 作者: Suparak S

Systematic mutagenesis of genes encoding predicted autotransported proteins of Burkholderia pseudomallei identifies factors mediating virulence in mice, net intracellular replication and a novel protein conferring serum resistance.

• DOI: 10.1371/journal.pone.0121271
• 发表时间: 2015
• 期刊: PloS one
• 影响因子: 3.7
• 作者: Lazar Adler NR;Stevens MP;Dean RE;Saint RJ;Pankhania D;Prior JL;Atkins TP;Kessler B;Nithichanon A;Lertmemongkolchai G;Galyov EE
• 通讯作者: Galyov EE

Identification of a predicted trimeric autotransporter adhesin required for biofilm formation of Burkholderia pseudomallei.

• DOI: 10.1371/journal.pone.0079461
• 发表时间: 2013
• 期刊: PloS one
• 影响因子: 3.7
• 作者: Lazar Adler NR;Dean RE;Saint RJ;Stevens MP;Prior JL;Atkins TP;Galyov EE
• 通讯作者: Galyov EE