Identification of factors required for Listeria motility

鉴定李斯特菌运动所需的因素

• 批准号: 6445269
• 负责人: ROBERT L JENG
• 金额: $ 3.83万
• 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/6445269
• 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 涉及运动的细胞行为范围。许多细菌病原体 作为其感染周期的一部分，劫持了宿主细胞细胞骨架。李斯特菌 例如，单核细胞增生蛋白聚合使肌动蛋白形成类似彗星的尾巴，这些尾巴 通过细胞质推动它。因为这些病原体绕过了许多 正常的细胞骨架对照，并且由于其运动已概括 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，我们描述了李斯特菌的使用来识别新因素 参与肌动蛋白动力学和力产生。我们发现李斯特菌 在不同细胞提取物中以不同的速率移动。李斯特菌在高处移动 人血小板和青蛙鸡蛋提取物的速率，但小鼠的速率低或 牛脑提取物。在大脑提取物中添加高动力提取物 赋予大脑提取的快速运动。我们建议将此活动用于 纯化的成分存在于血小板和鸡蛋提取物中 快速细菌运动。具体来说，我们计划：（1）进一步表征 不同细胞提取物中的李斯特菌运动，（2）分级提取物和 遵循高动力活性以净化增强细菌的因素 运动，（3）识别和表征纯化的蛋白质。我们相信 这种方法将隔离肌动蛋白动力学和力涉及的新因素 一代。