Erwinia Pectic Enzyme Out System: A Model for Studying Exoprotein Secretion by Gram-Negative Bacteria

欧文氏菌果胶酶输出系统：研究革兰氏阴性菌外蛋白分泌的模型

• 批准号: 9106431
• 负责人: Alan Collmer
• 金额: --
• 依托单位: Cornell Univ - State: AWDS MADE PRIOR MAY 2010
• 依托单位国家: 美国
• 项目类别: Continuing grant
• 财政年份: 1991
• 资助国家: 美国
• 起止时间: 1991-08-15 至 1994-07-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=9106431&HistoricalAwards=false
• 关键词: Erwinia; Pectic; Enzyme; Out; System

The exoprotein secretion ("Out") system of the plant pathogens Erwinia chrysanthemi and E. carotovora provide a promising experimental model for studying the translocation of proteins across the two membranes that envelope gram-negative bacteria. The bacteria selectively secrete a remarkable array of pectic enzymes and other exoproteins. A cluster of E. chrysanthemi "out" genes enables Escherichia coli to secrete E. chrysanthemi exoproteins. The E. chrysanthemi "out" genes are homologous with the pullulanase secretion genes of Klebsiella pneumoniae, suggesting that the Erwinia model will be broadly relevant. The two Erwinia "out" systems are conveniently specific for their respective exoproteins. The objectives of this project are to complete the characterization of the E. chrysanthemi "out" genes and encoded proteins by DNA sequence analysis and Tn5tac1 mutagenesis, identify the E. carotovora and E. chrysanthemi "out" components controlling exoprotein secretion specificity, and explore the secretion ability of hybrid proteins and deletion derivatives involving E. chrysanthemi PelE and either E. carotovora Pel1 or beta-lactamase. An adequate understanding of the "out" pathway could lead to general techniques for either disrupting the secretion of virulence proteins or enhancing secretion of beneficial proteins.

植物病原体菊欧文氏菌和胡萝卜软腐菌的外蛋白分泌（“Out”）系统为研究蛋白质跨包膜革兰氏阴性菌的两个膜的易位提供了一个有前景的实验模型。这些细菌选择性地分泌一系列显着的果胶酶和其他外蛋白。菊花大肠杆菌“out”基因簇使大肠杆菌能够分泌菊花大肠杆菌外蛋白。 E. chrysanthemi“out”基因与肺炎克雷伯菌的支链淀粉酶分泌基因同源，表明欧文氏菌模型具有广泛的相关性。两个欧文氏菌“输出”系统对其各自的外蛋白很方便地具有特异性。本项目的目标是通过DNA序列分析和Tn5tac1诱变完成菊花“out”基因和编码蛋白的表征，鉴定胡萝卜软叶菊和菊花“out”控制外蛋白分泌特异性的成分，并探索涉及菊花 E. chrysanthemi PelE 和 E. carotovora Pel1 或 β-内酰胺酶的杂合蛋白和缺失衍生物的分泌能力。对“外出”途径的充分了解可能会导致开发出破坏毒力蛋白分泌或增强有益蛋白分泌的通用技术。