CAMPYLOBACTER JEJUNI CYTOLETHAL DISTENDING TOXIN

空肠弯曲菌细胞致死膨胀毒素

• 批准号: 6226259
• 负责人: CAROL L PICKETT
• 金额: $ 28.96万
• 依托单位: UNIVERSITY OF KENTUCKY
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-09-30 至 2002-09-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6226259
• 关键词: Guillain Barre syndrome; Saccharomyces cerevisiae; bacteria infection mechanism; bacterial cytopathogenic effect; bacterial genetics; bacterial toxicology; bacterial toxins; cell cycle; cytotoxicity; fungal genetics; gastroenteritis; gene expression; gram negative bacteria; host organism interaction; intermolecular interaction; molecular site; protein structure function; site directed mutagenesis; tissue /cell culture; yeast two hybrid system

DESCRIPTION (Adapted from the Applicant's Abstract): Campylobacter jejuni is currently the most common bacterial cause of diarrheal disease in humans in the United States. It has also recently been strongly implicated as one of the most common infections to precede the development of Guillain-Barre syndrome. Thus far, only one C. jejuni toxin, cytolethal distending toxin (CDT), has been definitively proven to exist. Recent work has shown that the C. jejuni CDT is a member of a new family of toxins that can cause a G2 cell cycle block in certain mammalian cultured cells. However, little is known about the specific functions of this toxin and how it contributes to C. jejuni disease. The long-term goals of this proposal are to understand how CDT causes a cell cycle block and to relate this understanding to disease. In particular, the investigator plans to examine the functional roles of 3 CDT proteins (CdtA, CdtB, and CdtC) in causing the G2 block. The proposal seeks to elucidate the target(s) of CDT in sensitive cells, and determine the roles of the Cdt proteins in causing the toxic effect. The specific aims of the proposal are to test the hypothesis that one or more of the Cdt proteins function to bring about the G2 block in various types of eucaryotic cells. In Aim 1, a variety of specific mutations will be introduced into each of the Cdt proteins. The effects of the mutations on toxicity and on selected other properties will be tested. In Aim 2, yeast 2-hybrid techniques will be used to identify HeLa cell proteins that interact with Cdt proteins. In Aim 3, a yeast expression system will be exploited to identify how Cdt causes the G2 block. In Aim 4, a HeLa cell expression system will be used to complement and extend the studies in the first 3 aims. In summary, these studies are designed to increase our understanding of CDT and the mechanism by which it causes a G2 block.

描述（改编自申请人的摘要）：空肠弯曲菌是 目前，这是人类腹泻病最常见的细菌原因 美国。最近，它也被强烈暗示为最严重的问题之一。 发生格林-巴利综合征之前的常见感染。因此 迄今为止，只有一种空肠弯曲菌毒素，细胞致死膨胀毒素（CDT），已被 明确证明存在。最近的工作表明，空肠弯曲菌 CDT 是 一种新的毒素家族的成员，可导致 G2 细胞周期阻滞 某些哺乳动物培养细胞。但具体情况却鲜为人知 这种毒素的功能以及它如何导致空肠弯曲菌疾病。 该提案的长期目标是了解 CDT 如何导致细胞 循环阻滞并将这种理解与疾病联系起来。特别是， 研究人员计划检查 3 种 CDT 蛋白（CdtA、 CdtB 和 CdtC) 导致 G2 阻滞。该提案旨在阐明 敏感细胞中 CDT 的靶点，并确定 Cdt 的作用 蛋白质引起毒性作用。该提案的具体目标是 检验一种或多种 Cdt 蛋白的功能假设 关于各种类型真核细胞中的 G2 阻滞。在目标 1 中，各种 特定的突变将被引入到每个 Cdt 蛋白中。这 突变对毒性和选定的其他特性的影响将是 已测试。在目标 2 中，酵母 2 杂交技术将用于鉴定 HeLa 细胞 与 Cdt 蛋白相互作用的蛋白质。目标 3，酵母表达系统 将被用来识别 Cdt 如何导致 G2 阻塞。在目标 4 中，海拉 细胞表达系统将用于补充和扩展该领域的研究 前 3 个目标。总之，这些研究旨在提高我们的 了解 CDT 及其导致 G2 阻滞的机制。