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Evolution and consequences of multidrug resistant ribosome

多重耐药核糖体的进化和后果

基本信息

批准号：
10673677
负责人：
Mee-Ngan F Yap
金额：
$ 46.34万
依托单位：
NORTHWESTERN UNIVERSITY AT CHICAGO
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-09-16 至 2025-08-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10673677
关键词：
Address Adenine Affect Antibiotics Antimicrobial Resistance Bacterial Infections Binding Biochemical Biochemistry Biological Assay Carotenoids Catalysis Cells Cessation of life Ciprofloxacin Clinical Communities Complement Complex Coupled Cryoelectron Microscopy Crystallization DNA Gyrase Data Disulfides Enzymatic Biochemistry Enzymes Evolution Exhibits Family Gene Expression Generations Genes Genetic Polymorphism Genetic Transcription Genome Goals Gram-Positive Bacteria Growth Health Care Costs Impairment In Vitro Infection Knowledge Laboratories Link Macrolides Maps Mass Spectrum Analysis Measures Mediating Medical Messenger RNA Methylation Methyltransferase Methyltransferase Gene Mining Modeling Modification Molecular Multi-Drug Resistance Mutation Nucleotides Operon Pathway interactions Peptides Population Production Protein Biosynthesis Protein Inhibition Proteome Proteomics RNA RNA Binding Regulatory Pathway Reporter Resistance Ribosomal RNA Ribosomes Role Site Site-Directed Mutagenesis Staphylococcus aureus Streptogramins Structure System Testing Therapeutic Toll-like receptors Translations Treatment Failure Up-Regulation antimicrobial bacterial genetics bacterial resistance comparative genomics constitutive expression cost crosslink cytotoxic design experimental study fitness gel mobility shift assay genetic approach genome sequencing global health human disease human pathogen improved in vivo inhibitor innovation insight lincosamide next generation sequencing novel strategies posttranscriptional preference public health relevance resistance mechanism ribosome profiling structural biology trait translatome virulence gene

项目摘要

ABSTRACT Posttranscriptional modifications of bacterial and eukaryotic ribosomes are linked to many human diseases, but the precise role of most modifications remains undefined. Dimethylation of a universally conserved adenine, A2058, in bacterial rRNA causes cross-resistance against all three critically important families of antibiotics (macrolides, lincosamides, and streptogramins (MLS)). A2058 dimethylation occludes MLS from the ribosome, thereby allowing normal protein biosynthesis and bacterial growth. The thirty-five classes of Erm methyltransferases responsible for A2058 dimethylation are invariantly encoded by a two-gene operon preceded by a short ribosome stalling leader sequence. These short stalling peptides considerably vary in size and sequence composition. The functional and evolutionary connections between the stalling sequence and its cognate erm gene are poorly understood. A previous `ribosome stalling' model suggests that macrolide-mediated translational stalling of the leader sequence is required for the upregulation of downstream co-transcribed erm, but clinical surveillance and our data indicate the existence of an alternative pathway. Our unpublished data further show that collateral sensitivity to unrelated antibiotics, reduction in virulence gene expression, accumulation of inactive ribosomes, and loss of in vivo fitness are all part of the trade-offs associated with the A2058 dimethylated ribosome. The exact mechanistic links between these traits are unknown. There is also an unmet need to understand the mechanism by which Erm recognizes and acts on 23S rRNA. This proposal will use a multi-pronged approach consisting of high-precision next-generation sequencing, bacterial genetics, proteomics, comparative genomics, biochemistry and structural biology to address three central questions: What are the underlying mechanisms of the trade-offs conferred by the A2058 dimethylated ribosome? How does the erm operon evolve, and how is the expression of erm regulated? How does Erm find its target substrate RNA? The erm operons are widespread among nosocomial Gram-negative and Gram-positive bacteria, addressing these questions will offer significant mechanistic insight into new antimicrobial strategies tailored to disrupt these biochemical interactions and regulatory pathways.

摘要