Role of Non-Coding RNAs in P. aeruginosa Biofilm Development

非编码 RNA 在铜绿假单胞菌生物膜发育中的作用

• 批准号: 7643474
• 负责人: MICHAEL J FRANKLIN
• 金额: $ 17.81万
• 依托单位: MONTANA STATE UNIVERSITY - BOZEMAN
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-06-25 至 2011-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7643474
• 关键词: 5' Untranslated Regions; Antibiotic Resistance; Antibiotic Therapy; Bacteria sigma factor KatF protein; Bioinformatics; Biological; Cell physiology; Code; Custom; Cystic Fibrosis; Development; Functional RNA; Genes; Genome; Genome Mappings; Goals; Growth; Hereditary Disease; Infection; Intercistronic Region; Iron; Lung; Messenger RNA; Microbial Biofilms; Molecular Target; Northern Blotting; Pathogenesis; Patients; Phase; Positioning Attribute; Process; Production; Proteins; Proteomics; Pseudomonas aeruginosa; Research Personnel; Role; Tissues; Virulence Factors; base; design; novel; pathogen; public health relevance; quorum sensing; research study

DESCRIPTION (provided by applicant): Pseudomonas aeruginosa is an opportunistic pathogen that forms biofilm-associated infections on the pulmonary secretions of patients with the genetic disorder cystic fibrosis (CF). These infections are often impossible to eradicate with traditional antibiotic therapies. Therefore, investigators have performed global transcriptional and proteomics studies on P. aeruginosa to identify new molecular targets that are induced in P. aeruginosa during infectious processes. In many cases, production of specific proteins is regulated at the post-transcriptional level by small trans-acting RNAs or by cis-acting RNAs associated with the 5'-untranslated region of the mRNA. These non-protein-coding RNAs (ncRNAs) control expression of many cellular processes, including those involved in quorum sensing and virulence factor production. Approximately twenty P. aeruginosa ncRNAs have been identified previously. Based on our bioinformatics studies, this number is only a fraction of those actually encoded by P. aeruginosa. We predict that over 100 ncRNAs are encoded in the intergenic regions of the P. aeruginosa genome, and that others may be misannotated as protein-coding sequence. We have demonstrated expression of many of these predicted ncRNAs by using custom-designed microarrays and by Northern blotting experiments. Our present goals are to: (I) Map the genome position of the P. aeruginosa ncRNA, and to represent each on a custom microarray with tiled probes. Expression of these ncRNAs will be determined by using microarrays, Northern blotting, and mutational studies. (II) Define the biological activities of a novel ncRNA, located between P. aeruginosa genes PA4634 and PA4635. This small ncRNA is expressed only during stationary phase when P. aeruginosa is starved for iron, conditions likely to exist in the interiors of infectious biofilms. We will characterize the expression of this ncRNA and identify its mRNA targets. Our long-term goal is to identify new molecular targets that can be used for anti-P. aeruginosa therapies. These studies will facilitate this goal by identifying and characterizing many new regulatory molecules important in P. aeruginosa pathogenesis. PUBLIC HEALTH RELEVANCE: The opportunistic pathogen, Pseudomonas aeruginosa, colonizes pulmonary tissue of patients with the genetic disorder, cystic fibrosis. These infections are often resistant to antibiotic treatment. The goals of this study are to define post- transcriptional regulatory processes that occur during P. aeruginosa infection, in order to define novel molecular targets for anti-P. aeruginosa therapies.

描述（由申请人提供）：铜绿假单胞菌是一种机会性病原体，可在患有遗传性疾病囊性纤维化（CF）的患者的肺部分泌物上形成生物膜相关感染。这些感染通常无法用传统抗生素疗法根除。因此，研究人员对铜绿假单胞菌进行了全局转录和蛋白质组学研究，以确定铜绿假单胞菌在感染过程中诱导的新分子靶标。在许多情况下，特定蛋白质的产生在转录后水平受到小反式作用RNA或与mRNA 5'-非翻译区相关的顺式作用RNA的调节。这些非蛋白编码 RNA (ncRNA) 控制许多细胞过程的表达，包括参与群体感应和毒力因子产生的过程。先前已鉴定出大约二十种铜绿假单胞菌 ncRNA。根据我们的生物信息学研究，这个数字只是铜绿假单胞菌实际编码的一小部分。我们预测超过 100 个 ncRNA 被编码在铜绿假单胞菌基因组的基因间区域中，而其他的可能被错误地注释为蛋白质编码序列。我们通过使用定制设计的微阵列和 Northern 印迹实验证明了许多这些预测的 ncRNA 的表达。我们目前的目标是： (I) 绘制铜绿假单胞菌 ncRNA 的基因组位置图，并用平铺探针在定制微阵列上表示每个位置。这些 ncRNA 的表达将通过微阵列、Northern 印迹和突变研究来确定。 (II) 定义位于铜绿假单胞菌基因 PA4634 和 PA4635 之间的新型 ncRNA 的生物活性。这种小 ncRNA 仅在铜绿假单胞菌缺铁的稳定期表达，这种情况可能存在于感染性生物膜的内部。我们将表征该 ncRNA 的表达并确定其 mRNA 靶标。我们的长期目标是确定可用于抗 P 的新分子靶点。铜绿假单胞菌疗法。这些研究将通过鉴定和表征许多在铜绿假单胞菌发病机制中重要的新调节分子来促进这一目标的实现。 公共卫生相关性：机会性病原体铜绿假单胞菌定植于患有遗传性疾病囊性纤维化的患者的肺组织中。这些感染通常对抗生素治疗具有耐药性。本研究的目的是确定铜绿假单胞菌感染期间发生的转录后调控过程，从而确定抗铜绿假单胞菌的新分子靶标。铜绿假单胞菌疗法。