POST-TRANSCRIPTIONAL RRNA MODIFICATION FOR THE 30S RIBOSOMAL SUBUNIT

30S 核糖体亚基的转录后 RRNA 修饰

• 批准号: 8169319
• 负责人: Gerwald Jogl
• 金额: $ 0.35万
• 依托单位: CORNELL UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-04-01 至 2011-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8169319
• 关键词: Antibiotic Resistance; Antibiotics; Bacteria; Binding; Collaborations; Complex; Computer Retrieval of Information on Scientific Projects Database; DNA Sequence Rearrangement; Funding; Grant; Institution; Lead; Modification; Mutation; Organism; Post-Translational Protein Processing; Proteins; Research; Research Personnel; Resources; Ribosomal RNA; Ribosomes; Source; Streptomycin; Structure; United States National Institutes of Health; Universities; mutant; rRNA Genes

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Post-transcriptional and post-translational modification of ribosomal RNA and proteins is common in most or-ganisms, but the function of many of these modifications is not known. In bacteria, loss of some modifications leads to increased or decreased resistance to antibiotic drugs. We investigate the significance of these modi-fications for ribosome structure and function. This project is carried out in collaboration with Albert Dahlberg at Brown University, Venki Ramakrishnan at the MRC, UK, and Frank Murphy at the APS. We have now produced crystals for 30S ribosomal subunits from T. thermophilus with the gene for the rRNA methyltrans-ferase KsgA deleted. The KsgA dimethyltransferase modifies residues A1518 and A1519 of the 16S rRNA. This modification appears to be conserved in all organisms and confers sensitivity to the antibiotic kasu-gamycin. Structural information for the unmodified 30S subunits will lead to a better understanding of the function of these modifications. In a second approach, we are studying the impact of mutations on ribosome structure. We have produced 30S subunit crystals for two mutant forms: the streptomycin-dependent mutant G524U and the reverting double mutant G524U / A10G. We plan to determine structures of the mutant 30S ribosomal subunits in the apo-form and in complex with streptomycin to understand the rearrangements that lead to the altered binding of the antibiotic in the mutant forms.

该副本是利用众多研究子项目之一 由NIH/NCRR资助的中心赠款提供的资源。子弹和 调查员（PI）可能已经从其他NIH来源获得了主要资金， 因此可以在其他清晰的条目中代表。列出的机构是 对于中心，这不一定是调查员的机构。 核糖体RNA和蛋白质的转录后和翻译后修饰在大多数有机物中都是常见的，但是许多这些修饰的功能尚不清楚。在细菌中，某些修饰的丧失导致对抗生素药物的耐药性升高或降低。我们研究了这些修饰问题对核糖体结构和功能的重要性。该项目与布朗大学，英国MRC的Venki Ramakrishnan的Albert Dahlberg和APS的Frank Murphy合作进行。现在，我们已经从T. thermothilus的30S核糖体亚基中生产了rRNA甲基trans-屈服酶KSGA的基因的晶体。 KSGA二甲基转移酶修饰16S rRNA的残基A1518和A1519。这种修饰似乎在所有生物体中都是保守的，并且赋予对抗生素kasu-gamycin的敏感性。未修改的30S亚基的结构信息将使人们更好地理解这些修饰的功能。 在第二种方法中，我们正在研究突变对核糖体结构的影响。我们为两种突变形式生产了30 s亚基晶体：链霉素依赖性突变体G524U和恢复双突变体G524U / A10G。我们计划确定APO形式中突变体30S核糖体亚基的结构，并与链霉素复杂化，以了解会导致突变体形式中抗生素的结合改变的重排。