RNA in Viral DNA Packaging

病毒 DNA 包装中的 RNA

• 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
• 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

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碱基prohead RNA（PRNA）是分子运动的重要组成部分，将病毒基因组DNA包装到病毒前体帽（Prohead）中。该电动机是已知的最强分子电动机之一，产生大于60 pn的力。 O29是用于DNA包装的主要模型系统，是某些动物病毒（例如疱疹和腺病毒）中类似过程的模型。 PRNA通过在相同的分子之间通过分子间碱基对形成新型的环状五聚体来与prohead结合。然后，RNA环提供了一个支架，包装ATPase将其组装成其功能环形状。除了提供脚手架功能外，PRNA还可以在更高级的包装过程中（例如电动机内的协调和通信）发挥作用。研究该RNA依赖性DNA包装电机的结构和功能可能在发现抗病毒剂的靶标方面具有一般意义。另外，正在为纳米医学应用利用PRNA的性质。该研究的最终目标是确定PRNA在DNA易位机理中的结构和功能作用。在这里，我们采用高度综合的多学科方法，包括遗传和生化分析，X射线晶体学，冷冻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射线晶体学将用于通过PRNA和PRNA复杂形成的原子化来完成X射线晶体学，以完成PRNA的结构。 2）表征与位置定向诱变和嵌合RNA相关的PRNA的功能元素； 3）使用含有野生型和突变体PRNA的有序的杂体PRNA环研究PRNA介导的组装和运动通信和协调。

项目成果

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)
• 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

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.