古菌 sRNP 的结构和功能分析-猫眼课题宝

古菌 sRNP 的结构和功能分析

批准号：

7265306

负责人：

BERNARD ANDREW BROWN

金额：

$24.51万

依托单位：

NORTH CAROLINA STATE UNIVERSITY RALEIGH

依托单位国家：

美国

项目类别：

财政年份：

2005

资助国家：

美国

项目状态：

已结题

起止时间：

2005-08-01 至 2010-03-31

来源：

https://reporter.nih.gov/project-details/7265306

关键词：

Accounting Acids Active Sites Address Adopted Aeropyrum Affect Affinity Affinity Chromatography Amino Acid Motifs Amino Acids Anabolism Archaea Archaeal Proteins Archaeoglobus fulgidus Architecture Area Arginine Back Base Pairing Base Sequence Binding Binding Sites Biochemical Biochemical Genetics Biogenesis Biohazardous Substance Biological Biological Assay Biological Process Blood capillaries Bone Marrow Diseases Boxing Bromine Bromouridine Buffaloes Buffers C-terminal Caring Catalysis Catalytic RNA Cell Nucleolus Cell Nucleus Cell Survival Cells Centrifugation Characteristics Charge Chelating Agents Chemicals Chromatography Circular Dichroism Circular Dichroism Spectroscopy Classification Cleaved cell Cobalt Coiled-Coil Domain Collaborations Color Communication Complement Complement Factor B Complementary RNA Complex Condition Consensus Sequence Consultations Core Protein Coupled Crystallization Crystallography Cytidine Cytolysis Data Data Collection Databases Decompression Sickness Deposition Detergents Development Dialysis procedure Diffusion Dimensions Dimerization Disease Disruption Dissection Divalent Cations Drops Dyskeratosis Congenita Edetic Acid Electrophoretic Mobility Shift Assay Electrostatics Elements Ensure Environment Enzymes Equilibrium Equipment Escherichia coli Essential Genes Etiology Eukaryota Eukaryotic Cell Evaluation Event Evolution Exhibits Experimental Designs Exposure to Factor Analysis Failure Family Figs - dietary Florida Fluorescence Fluorescence Resonance Energy Transfer Fluorescence Spectroscopy Foundations Freedom Funding Future Gel Gel Chromatography Genes Genetic Genetic Transcription Goals Gray unit of radiation dose Growth Guide RNA Haloarcula marismortui Hand Hazardous Substances Health Hearing Heavy Metals Helix (Snails)Helix-Loop-Helix Motifs Hepatitis Delta Virus Hepatocyte Growth Factor Heterodimerization Home environment Homodimerization Homologous Gene Homologous Protein Housing Human Hydrogen Bonding Hydrolysis Hydroxyl Radical Imagery Imidazole In Vitro Incubated Individual Institutes Instruction Investigation Iodine Ion-Exchange Chromatography Procedure Ionizing radiation Ions Iridium Knock-out Label Laboratories Laboratory Finding Lasers Lead Left Length Lesion Letters Leucine Life Ligands Light Link Lysine Macromolecular Complexes Magnesium Magnesium Chloride Malignant Neoplasms Mammals Manuals Manufacturer Name Maps Mass Spectrum Analysis Memorial Sloan-Kettering Cancer Center Mercury Metals Methanococcus Methionine Methodology Methods Methylation Methyltransferase Minor Modeling Modification Molecular Molecular Conformation Molecular Target Monitor Mothers Multienzyme Complexes Mus Muscle Rigidity Mutagenesis Mutate Mutation N-terminal Nickel Noise None or Not Applicable North Carolina Nucleic Acids Nucleolar Proteins Nucleosides Nucleotides Numbers Object Attachment Organism Osmium Pathogenesis Pattern Phase Physical Dialysis Placement Plant Resins Plant Roots Play Pliability Population Positioning Attribute Predisposition Preparation Principal Investigator Probability Procedures Process Production Productivity Property Protein Analysis Protein Binding Protein Biosynthesis Protein Overexpression Protein Splicing Proteins Proteolysis Protocols documentation Pseudouridine Publishing Pyrococcus R-factor RNA RNA Binding RNA Folding RNA Sequences RNA chemical synthesis RNA-Protein Interaction Radiolabeled Reaction Reagent Recombinant DNA Recombinants Regulation Relative (related person)Reporting Research Research Design Research Personnel Research Project Grants Resolution Resources Ribonucleases Ribonucleoproteins Ribonucleotides Ribose Ribosomal Proteins Ribosomal RNA Ribosomes Roentgen Rays Role Role playing therapy Rotation Route Running Safety Salts Sampling Saturn Scanning Scheme Screening procedure Selenomethionine Side Signal Recognition Particle Signal Transduction Simulate Site Site-Directed Mutagenesis Skin Small Nuclear RNA Small Nuclear Ribonucleoproteins Small Nucleolar RNA Small Nucleolar Ribonucleoproteins Small RNA Sodium Chloride Sodium Dodecyl Sulfate-PAGE Sodium Iodide Solid Solutions Solvents Source Specificity Spectrometry Spectrum Analysis Speed Spliceosomes Staging Standards of Weights and Measures Stem cells Structure Structure-Activity Relationship Sulfolobus solfataricus Surface Surveys Synchrotrons Syndrome System Techniques Telomerase Telomerase RNA Component Telomere Maintenance Telomere Shortening Temperature Tern Tertiary Protein Structure Testing Thermus thermophilus Thinking Thrombin Time Transcript Transfer RNA Translating Translations U1A protein U4 Small Nuclear Ribonucleoproteins U4 small nuclear RNA Uncertainty Universities Urea Uridine Variant Vendor Vertebral column Water Weight Work X-Ray Crystallography Yeasts Zinc absorption antitermination base beamline capillary cell dimension chelation crosslink day density design design and construction desire dimer distilled alcoholic beverage divalent metal ear helix electron density experience expression vector falls fast protein liquid chromatography fibrillarin follow-up forest gel electrophoresis human subject hydroxyl group improved indexing innovation insight instrument interest intermolecular interaction light scattering loss of function macromolecule metaplastic cell transformation mutant novel nuclease nucleic acid structure numb protein phosphorothioate pressure prevent programs prolylarginine protein crosslink protein protein interaction protein structure rRNA Precursor radiotracer radius bone structure reconstitution research study resistance factors restraint size sodium bromide solute statistics stem stoichiometry structural biology success sugar synchrotron radiation telomere trend tumor tumorigenic vapor wasting

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中文摘要

翻译

描述(申请人提供)：核糖体RNA在生物发生过程中被共价修饰。未能正确修饰rRNA会影响蛋白质合成的效率，并可能参与肿瘤的形成和疾病的发病机制。真核生物和古生物中最普遍的rRNA修饰之一是核糖2‘-羟基的2’-O-甲基化。这些修饰是由保守的盒C/D小核仁RNA家族指导的，它包含终端盒C/D和并列的内盒CO/D没有图案。目前，古菌盒C‘/D’小核糖核蛋白(SRNPs)是目前唯一可以在体外重组获得具有酶活性的复合体的RNA引导的核苷酸修饰系统。这些系统的结构研究已经成熟，可以确定复杂组装的机制及其与调控的联系，并评估突变如何影响甲基化活性。这项研究的长期目标是研究古生物中BOX C/D引导的2‘-O-甲基化系统的结构生物学，并将其与真核生物的相应系统进行比较。这项研究将专门研究简氏甲烷球菌和Pernix Aeropyrum box C/D rRNPs的结构和活性关系。在这些系统中，L7Ae核心蛋白通过结合box C/D RNA并与Nop56/58和纤维蛋白核心蛋白建立相互作用来启动sRNP组装。目的是阐明蛋白质-RNA和蛋白质/蛋白质的相互作用、结合、折叠和组装是如何调节生物活性的。具体目标是： 1.将单个和并列的古生物盒C/D和C‘/D’SRNA基序与孤立和并列的L7Ae-box C/D和L7Ae-C‘/D’sRNP复合体的结构进行比较和对比； 2.研究古生菌核心蛋白L7Ae在启动sRNP组装中的作用以及锌在这些相互作用中的可能作用。这项研究的基本原理是增加补充的生物物理和结构数据，以支持越来越多的关于古生物盒C/D RNA修饰系统的生化研究。这些研究将对核糖核蛋白酶复合体的进化和发展提供洞察，并阐明古生物和真核生物sRNP系统之间的异同。

英文摘要

DESCRIPTION (provided by applicant): Ribosomal RNAs are covalently modified during biogenesis. Failure to properly modify rRNA can affect the efficiency of protein synthesis and may be involved in tumor-formation and disease pathogenesis. One of the most prevalent rRNA modification in both eukaryotes and archaea is the 2'-O-methylation of ribose 2'-hydroxyl groups. These modifications are directed by the conserved family of box C/D small nucleolar RNAs, which contain terminal box C/D and juxtaposed internal box CO/DO motifs. At present, archaeal box C'/D' small ribonucleoproteins (sRNPs) are the only RNA-guided nucleotide modification systems that can be reconstituted in vitro to obtain enzymatically active complexes. These systems are ripe for structural investigations that determine the mechanism of complex-assembly and its linkage to regulation, and assess how mutations affect methylation activity. The long-term goals of this research are to investigate the structural biology of the box C/D-guided 2'-O-methylation systems in archaeal organisms and compare them to eukaryotic counterparts. This investigation will specifically examine the structure <--> activity relationship of the Methanocaldococcus jannaschii and Aeropyrum pernix box C/D sRNPs. In these systems, the L7Ae core protein initiates sRNP assembly by binding box C/D RNAs and establishing interactions with the Nop56/58 and fibrillarin core proteins. The goals are to elucidate how protein-RNA and protein/protein interactions, binding, folding, and assembly regulate biological activity. The specific aims are: 1. To compare and contrast the structures of the individual and juxtaposed archaeal box C/D and C'/D' sRNA motifs to those in isolated and juxtaposed L7Ae-box C/D and L7Ae-C'/D' sRNP complexes; and 2. To examine the archaeal core protein L7Ae's role in initiating sRNP assembly and the putative function of zinc in these interactions. The rationale for this investigation is to add complementary biophysical and structural data to support the mounting biochemical studies on archaeal box C/D RNA modification systems. These studies will provide insight to the evolution and development of ribonucleoprotein enzyme complexes, and illuminate the similarities and difference between the archaeal and eukaryotic sRNP systems.

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