STABILITY GENES OF E FAECALIS PLASMID PAD1

粪肠球菌质粒PAD1的稳定性基因

• 批准号: 6386664
• 负责人: KEITH E WEAVER
• 金额: $ 14.98万
• 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 2003-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6386664
• 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.

这项建议的广泛和长期目标是界定 PAR稳定性的分子作用机制和调节 粪肠球菌质粒pAD 1上编码的决定簇。par不 大于404个核苷酸，编码2个小RNA， 功能有证据表明，par的功能是作为一个后分离 杀戮系统两种RNA中较大的RNA 1是毒性组分 编码一个由33个氨基酸组成的多肽。较小的RNA RNA，RNAII，调节RNAi活性。据推测，RNAII 作为反义RNA抑制肽翻译，并且RNAi- 编码的肽抑制一些必需的宿主功能。四个具体 我们将致力于检验这一假设。第一，详细结构 将对par RNA及其相互作用复合物进行研究 在体外和体内。第二，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查。 第三，毒素靶标将通过鉴定宿主蛋白质来确定 与RNAi编码的肽相互作用。第四，其他质粒- 将确定与PAR相互作用的编码稳定性决定因素 通过用各种构建体进行竞争实验， 表征位于邻近的潜在活性分区系统， 段落人们希望平价制度可以以某种方式加以操纵， 为肠球菌感染提供有用的抗菌治疗。 或者，PAR毒素靶标的鉴定可以提供 有效的新抗生素设计的有用的目标， 肠球菌。