Identification of factors required for Listeria motility

鉴定李斯特菌运动所需的因素

• 批准号: 6622323
• 负责人: ROBERT L JENG
• 金额: $ 4.64万
• 依托单位: UNIVERSITY OF CALIFORNIA BERKELEY
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-02-01 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/6622323
• 关键词: Listeria; Listeria infections; Xenopus oocyte; actins; bacteria infection mechanism; bacterial genetics; bacterial proteins; cell component structure /function; cell free system; cell motility; chromatography; cytoskeletal proteins; cytoskeleton; fluorescence microscopy; host organism interaction; human tissue; intermolecular interaction; matrix assisted laser desorption ionization; method development; molecular cloning; protein purification; protein sequence; protein structure function; time resolved data; video microscopy

DESCRIPTION (provided by applicant): The actin cytoskeleton participates in a range of cellular behaviors involving motility. A number of bacterial pathogens hijack the host cell cytoskeleton as part of their infectious cycle. Listeria monocytogenes, for example, polymerizes actin to form comet-like tails that propels it through the cytoplasm. Because these pathogens bypass many of the normal cytoskeletal controls and because their movement has been recapitulated in cell-free extracts, they have been used as model systems to study actin polymerization. Here, we describe the use of Listeria to identify novel factors involved in actin dynamics and force generation. We have found that Listeria move at different rates in different cells extracts. Listeria move at high rates in human platelet and frog egg extracts, but at low rates in mouse or bovine brain extracts. Addition of high-motility extracts to the brain extracts confers fast movement to brain extract. We propose to use this activity to purify components present in platelet and egg extracts that are involved in fast bacterial motility. Specifically, we plan to: (1) further characterize Listeria motility in different cell extracts, (2) fractionate extracts and follow high-motility activity to purify factors that enhance bacterial movement, and (3) identify and characterize the purified proteins. We believe this approach will isolate novel factors involved in actin dynamics and force generation.

描述（由申请人提供）：肌动蛋白细胞骨架参与 涉及运动的一系列细胞行为。多种细菌病原体 劫持宿主细胞的细胞骨架作为其感染周期的一部分。李斯特菌 例如，单增细胞将肌动蛋白聚合形成彗星状尾巴， 推动它穿过细胞质。因为这些病原体绕过了许多 正常的细胞骨架控制，因为它们的运动已被重现 在无细胞提取物中，它们已被用作研究肌动蛋白的模型系统 聚合。在这里，我们描述了使用李斯特菌来识别新因素 参与肌动蛋白动力学和力的产生。我们发现李斯特菌 在不同的细胞提取物中以不同的速率移动。李斯特菌高位移动 人类血小板和青蛙卵提取物中的比率，但在小鼠或小鼠中的比率较低 牛脑提取物。在脑提取物中添加高活力提取物 赋予大脑提取物快速运动。我们建议利用这项活动 纯化血小板和鸡蛋提取物中涉及的成分 细菌运动快。具体来说，我们计划：（1）进一步表征 李斯特菌在不同细胞提取物中的运动性，(2) 分级提取物和 遵循高活力活性来纯化增强细菌的因子 运动，(3) 鉴定和表征纯化的蛋白质。我们相信 这种方法将分离出参与肌动蛋白动力学和力的新因素 一代。