RNA in Viral DNA Packaging

病毒 DNA 包装中的 RNA

基本信息

批准号：
9319766
负责人：
Paul James Jardine
金额：
$ 35.92万
依托单位：
UNIVERSITY OF MINNESOTA
依托单位国家：
美国
项目类别：
财政年份：
1999
资助国家：
美国
起止时间：
1999-09-01 至 2020-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9319766
关键词：
3-Dimensional ATP Hydrolysis ATP phosphohydrolase ATPase Domain Address Adenoviruses Animals Antiviral Agents Bacillus subtilis Bacteriophages Base Pairing Binding Binding Sites Biochemical Biological Biological Models Capsid Communication Complex Cryoelectron Microscopy DNA DNA Packaging Defect Development Double Stranded DNA Virus Electrostatics Elements Entropy Eukaryotic Cell Event Genetic Genome Genomic DNA Goals Head Herpesviridae Individual Infection Length Link Mediating Medical Modeling Molecular Machines Molecular Motors Motor Mutagenesis Nature Nucleic Acids Periodicity Pharmaceutical Preparations Play Positioning Attribute Process Property Proteins RNA RNA Binding RNA Phages Research Roentgen Rays Role Shapes Site-Directed Mutagenesis Specificity Structure Structure-Activity Relationship Variant Viral Virus Virus Assembly Virus Diseases Work X-Ray Crystallography alpha helix base biological systems crosslink crystallinity design ds-DNA experimental study insight interdisciplinary approach laser tweezer mutant nanomedicine novel novel therapeutics operation pressure public health relevance reconstruction scaffold single molecule targeted treatment viral DNA

项目摘要

DESCRIPTION (provided by applicant): The 174-base prohead RNA (pRNA) of bacteriophage o29 is an essential component of the molecular motor that packages the viral genomic DNA into the viral precursor capsid (prohead). This motor is one of the strongest molecular motors known, generating forces greater than 60 pN. o29 is a premier model system for DNA packaging and serves as a model for analogous processes in certain animal viruses, such as herpes- and adenoviruses. pRNA binds to the prohead by forming a novel cyclic pentamer via intermolecular base pairing between identical molecules. The RNA ring then provides a scaffold upon which the packaging ATPase assembles into its functional ring shape. In addition to providing a scaffold function, pRNA is hypothesized to function in higher order processes of packaging, such as coordination and communication within the motor. Study of the structure and function of this RNA-dependent DNA packaging motor may have general significance in uncovering targets for antiviral agents. Additionally, the properties of pRNA are being exploited for nanomedicine applications. The ultimate goal of the research is to determine the structure and functional roles of pRNA in the mechanism of DNA translocation. Here we employ a highly integrated, multi-disciplinary approach, including genetic and biochemical analysis, X-ray crystallography, cryoEM 3-D reconstruction and single molecule laser tweezers to investigate the structure/function relationship of pRNA: 1) X-ray crystallography will be used to complete the atomic structure of pRNA, and pRNA- ATPase complex formation will be characterized by biochemical analysis; 2) characterize functional elements of pRNA involved in motor assembly, communication and coordination by site-directed mutagenesis and chimeric RNAs; and 3) investigate pRNA-mediated assembly and motor communication and coordination using ordered heteromeric pRNA rings containing wild-type and mutant pRNAs.

描述（由申请人提供）：噬菌体 o29 的 174 个碱基的前头 RNA (pRNA) 是将病毒基因组 DNA 包装到病毒前体衣壳（前头）中的分子马达的重要组成部分。该电机是已知最强的分子电机之一，产生的力大于 60 pN。 o29 是 DNA 包装的首要模型系统，可作为某些动物病毒（例如疱疹病毒和腺病毒）中类似过程的模型。 pRNA 通过相同分子之间的分子间碱基配对形成新型环状五聚体，从而与前头结合。然后，RNA 环提供了一个支架，包装 ATP 酶在该支架上组装成其功能性环形。除了提供支架功能外，pRNA 还被假设在更高阶的包装过程中发挥作用，例如电机内的协调和通信。研究这种依赖于 RNA 的 DNA 包装马达的结构和功能对于揭示抗病毒药物的靶点可能具有普遍意义。此外，pRNA 的特性也被用于纳米医学应用。研究的最终目标是确定pRNA在DNA易位机制中的结构和功能作用。在这里，我们采用高度集成的多学科方法，包括遗传和生化分析、X 射线晶体学、cryoEM 3-D 重建和单分子激光镊子来研究 pRNA 的结构/功能关系：1）X 射线晶体学将用于完成pRNA的原子结构，并通过生化分析来表征pRNA-ATP酶复合物的形成； 2) 通过定点诱变和嵌合 RNA 表征参与运动组装、通讯和协调的 pRNA 功能元件； 3) 使用含有野生型和突变型 pRNA 的有序异聚 pRNA 环研究 pRNA 介导的组装以及运动通讯和协调。

项目成果

期刊论文数量（23）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Role of the CCA bulge of prohead RNA of bacteriophage ø29 in DNA packaging.

噬菌体 29 前头 RNA 的 CCA 凸起在 DNA 包装中的作用。

DOI：
10.1016/j.jmb.2008.08.056
发表时间：
2008
期刊：
Journal of molecular biology
影响因子：
5.6
作者：
Zhao,Wei;Morais,MarcC;Anderson,DwightL;Jardine,PaulJ;Grimes,Shelley
通讯作者：
Grimes,Shelley

Cas5d protein processes pre-crRNA and assembles into a cascade-like interference complex in subtype I-C/Dvulg CRISPR-Cas system.

DOI：
10.1016/j.str.2012.06.016
发表时间：
2012-09-05
期刊：
Structure (London, England : 1993)
影响因子：
0
作者：
Nam KH;Haitjema C;Liu X;Ding F;Wang H;DeLisa MP;Ke A
通讯作者：
Ke A

Substrate interactions and promiscuity in a viral DNA packaging motor.

DOI：
10.1038/nature08443
发表时间：
2009-10-01
期刊：
Nature
影响因子：
64.8
作者：
通讯作者：

Structural and Molecular Basis for Coordination in a Viral DNA Packaging Motor.

DOI：
10.1016/j.celrep.2016.01.058
发表时间：
2016-03-01
期刊：
Cell reports
影响因子：
8.8
作者：
Mao H;Saha M;Reyes-Aldrete E;Sherman MB;Woodson M;Atz R;Grimes S;Jardine PJ;Morais MC
通讯作者：
Morais MC

Defining molecular and domain boundaries in the bacteriophage phi29 DNA packaging motor.

DOI：
10.1016/j.str.2008.05.010
发表时间：
2008-08-06
期刊：
STRUCTURE
影响因子：
5.7
作者：
Morais, Marc C.;Koti, Jaya S.;Bowman, Valorie D.;Reyes-Aldrete, Emilio;Anderson, Dwight L.;Rossmann, Michael G.
通讯作者：
Rossmann, Michael G.