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Erwinia chrysanthemum和E. carotovora的外蛋白分泌（Out）系统为研究蛋白质在革兰氏阴性菌的两层膜上的易位提供了一个有希望的实验模型。这种细菌选择性地分泌大量果胶酶和其他外蛋白。一组菊花E.“out”基因使大肠杆菌能够分泌菊花E.外蛋白。菊花的“out”基因与肺炎克雷伯菌的普鲁兰酶分泌基因同源，这表明Erwinia模型将具有广泛的相关性。这两个Erwinia“out”系统方便地针对各自的外蛋白进行特异性处理。本项目旨在通过DNA序列分析和Tn5tac1诱变完成对菊花E. carotovora“out”基因和编码蛋白的鉴定，鉴定出控制外蛋白分泌特异性的E. carotovora和E.菊花E. cartovora Pel1或β -内酰胺酶的杂交蛋白和缺失衍生物的分泌能力。对“输出”途径的充分理解可能会导致破坏毒性蛋白分泌或增强有益蛋白分泌的通用技术。