Determining bacterial RNA polymerase functionalities required for sigma factor specific escape from antibiotic action.

如以下文献所述，基础研究在针对可受益于额外基础知识的领域时，对社会和经济目标的贡献最大：1）纳尔逊，R.R.，2004.市场经济与科学公地。Res. Policy 33：455-471; 2）Rosenberg，Nathan，and Richard R.纳尔逊（1994），“美国大学和工业技术进步”，Res. Policy 23：323-348; 3）罗森伯格，内森（1982），“科学如何外生？见Inside the Black Box（NY：剑桥University Press），p. 141-159.在本提案中，我们试图确定细菌如何应对用于控制其在感染环境中生长的药剂。了解细菌对抗菌剂的相对敏感性对于管理牲畜和人类感染非常重要，并为管理感染和设计新的抗菌剂提供信息。抗菌剂的一个有效靶点是基因表达的中心机制，即解码DNA的RNA聚合酶（RNAP）。了解各种RNAP全酶对抗菌剂的相对敏感性的基础是重要的，并谈到了如何解释灵敏度的内在水平--具有实际意义的东西。该应用程序的动机是基础知识中的差距，目前存在的差距仅仅是因为RNAP状态的范围--通过使用不同的附属因子（称为sigma因子）来反映--在评估靶向细菌RNAP的抗微生物剂的功效时，通常不考虑这些因素。各种RNAP对抗菌剂的反应是重要的，并且说明了如何解释生物体或基因集的内在敏感性水平。新出现的证据表明，转录因子，如RNAP σ因子，可以是形成耐药基因组的一组因子的成员。这表明，细胞的基本组成可以赋予抗微生物剂显著的不敏感性，而不会引起特定耐药机制的作用。由于转录因子如RNAP σ因子可以是耐药基因组的成员，我们希望弄清楚这些因子如何赋予RNAP对抗菌剂的耐受性。这些结果有望扩大我们对靶向细菌RNAP的抗生素作用的了解，以及RNA如何发挥作用以获取DNA中的遗传信息。我们将使用我们建立的方法来检查DNA解码过程中的哪些步骤对抗生素的作用敏感或耐药，特别是那些正在成为抗菌治疗新候选药物的药物。

Basic research best contributes to social and economic goals when targeted to areas that can benefit from additional fundamental knowledge, as reported in: 1) Nelson, R.R., 2004. The market economy and the scientific commons. Res. Policy 33: 455-471; 2) Rosenberg, Nathan, and Richard R. Nelson (1994), "American Universities and Technical Advance in Industry," Res. Policy 23:323-348; 3) Rosenberg, Nathan (1982), "How Exogenous is Science?" in Inside the Black Box (NY: Cambridge University Press), p. 141-159. In this proposal we seek to establish how bacteria respond to the agents that are used to control their growth in infection settings.Knowing the basis of relative sensitivities of bacteria to antimicrobials is important for managing infection in live stock and in humans, and informs strategies to manage infection and design new antimcrobials. One validated target for antimicrobials is the central machinery of gene expression, the enzyme RNA polymerase (RNAP) that decodes the DNA. Knowing the basis of relative sensitivities of various RNAP holoenzymes to antimicrobials is important, and speaks to how the intrinsic levels of sensitivity may be accounted for - something of practical significance.The application was motivated by the gap in basic knowledge which currently exists simply because the range of RNAP states - reflected by the use of differing accessory factors called sigma factors - are not often taken into account when assessing the efficacies of antimicrobials that target bacterial RNAPs. The response of various RNAPs to antimicrobials is important, and speaks to how the intrinsic levels of sensitivity of an organism or gene set might be accounted for. Emerging evidence shows that transcription factors, such as RNAP sigma factors, can be members of a group of factors that form the resistome. Showing that is, that the basic make up of the cell that can confer significant insensitivity to an antimicrobial without invoking the action of specific resistance mechanisms. Since transcription factors such as RNAP sigma factors can be members of a resistome, we wish to work out how these factors confer upon RNAP tolerance to antimicrobials. Outcomes arising are expected to enlarge our knowledge of the action of antibiotics that target bacterial RNAP, as well as how the RNA functions to access the genetic information in DNA. We will use methods established by us to examine which steps in the DNA decoding process are sensitive or resistant to the actions of antibiotics, in particular those agents which are emerging as new candidates for antimicrobial therapies.