STABILITY GENES OF E FAECALIS PLASMID PAD1

粪肠球菌质粒PAD1的稳定性基因

• 批准号: 2848499
• 负责人: KEITH E WEAVER
• 金额: $ 14.55万
• 依托单位: UNIVERSITY OF SOUTH DAKOTA
• 依托单位国家: 美国
• 财政年份: 1999
• 资助国家: 美国
• 起止时间: 1999-05-01 至 2002-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2848499
• 关键词: Enterococcus; bacterial RNA; bacterial genetics; bacterial toxins; chemical stability; genetic regulation; genetic regulatory element; nucleic acid sequence; nucleic acid structure; pheromone; plasmids; protein protein interaction; replicon; structural biology; yeast two hybrid system

The broad, long-term objective of this proposal is to define the molecular mechanisms of action and regulation of the par stability determinant encoded on the Enterococcus faecalis plasmid pAD1. par is no larger than 404 nucleotides and encodes 2 small RNAs essential for its function. Evidence indicates that par functions as a post-segregational killing system. The larger of the 2 RNAs, RNA1, is the toxic component of the system and encodes and 33 amino acid peptide. The smaller RNA, RNA, RNAII, regulates RNAI activity. It is hypothesized that RNAII inhibits peptide translation as an antisense RNA and that the RNAI- encoded peptide inhibits some essential host function. Four specific aims will be pursued to test this hypothesis. First, detailed structural studies on both par RNAs and their interaction complex will be performed in vitro and in vivo. Second, potential autoregulatory effects on par RNA production will be examined by a variety of genetic approaches. Third, the toxin target will be determined by identifying host proteins that interact with the RNAI-encoded peptide. Fourth, other plasmid- encoded stability determinants that interact with par will be identified by conducting competition experiments with various constructs and by characterizing a potential active partition system located adjacent to par. It is hoped that the par system could be manipulated in some way to provide a useful antibacterial treatment for enterococci infection. Alternatively, identification of the par toxin target could provide a useful target for the design of new antibiotics effective against the enterococci.

这项提案的广泛、长期目标是定义 PAR稳定性的分子作用机制及调控 粪肠球菌质粒PAD1上编码的决定簇。PAR不是 长于404个核苷酸，编码2个对ITS必不可少的小RNA 功能。证据表明，PAR起到了后种族隔离的作用 杀戮系统。两个RNAs中较大的RNA1是有毒成分 编码33个氨基酸的多肽。RNA越小， RNA，RNAII，调节RNAi活性。据推测RNAII 抑制作为反义RNA的肽翻译，并且RNAi- 编码的多肽抑制了一些基本的宿主功能。四个具体的 我们将致力于验证这一假说。第一，详细的结构 将对PAR RNAs及其相互作用复合体进行研究 在体外和体内。第二，面值上潜在的自动调节效应 RNA的产生将通过各种遗传方法进行检测。 第三，毒素靶标将通过识别宿主蛋白来确定。 与RNAi编码的多肽相互作用。第四，其他质粒- 将识别与PAR相互作用的编码稳定性决定因素 通过使用各种结构进行竞争实验，并通过 表征位于相邻位置的潜在活动分区系统 标准杆。希望平价制度能够以某种方式被操纵，以 为肠球菌感染提供有效的抗菌治疗。 或者，对PAR毒素靶标的识别可以提供 为设计有效对抗疟疾的新抗生素提供有用的目标 肠球菌。