Assembly and Structure of Bacteriophage Lambda

噬菌体 Lambda 的组装和结构

• 批准号: 8827783
• 负责人: ROGER W HENDRIX
• 金额: $ 55.87万
• 依托单位: UNIVERSITY OF PITTSBURGH AT PITTSBURGH
• 依托单位国家: 美国
• 财政年份: 1977
• 资助国家: 美国
• 起止时间: 1977-12-01 至 2018-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8827783
• 关键词: Address; Amino Acid Sequence; Amino Acids; Archaea; Attention; Bacteriophage lambda; Bacteriophages; Binding; Biochemical; Biological; Biological Models; Biological Process; Capsid; Capsid Proteins; Complex; Cyanobacterium; Data; Genes; Genetic; Genome; Goals; Grant; Head; Health; Herpesviridae; Investigation; Iron; Learning; Length; Location; Marines; Measures; Mediating; Methods; Minor; Molecular Chaperones; Mutation; N-terminal; Peptide Hydrolases; Peptide Sequence Determination; Phenotype; Positioning Attribute; Process; Proteins; Relative (related person); Ribosomes; Role; Rudiviridae; Solid; Streptomyces; Structural Protein; Structure; Sulfolobus; Sulfur; Tail; Testing; Translating; Translations; Tube; Virion; Virus; genome sequencing; human disease; mutant; nanomachine; protein function; protein structure; research study; terminase

DESCRIPTION (provided by applicant): The main emphasis of this grant is to understand how a protein nanomachine is assembled from its parts, how it functions once assembled, and how the mechanisms of assembly support the nanomachine's biological functions. To address these questions we will use the capsid of bacteriophage HK97 and the tail of bacteriophage??? Both of these offer experimental access to centrally important aspects of biological assembly: The Delta domain of the HK97 capsid protein is an assembly chaperone that mediates assembly of capsomers into capsid and also binds the protease and brings it into the inside of the capsid. It also has a role in getting the portal into position and probably has a crucial role in nucleating capsid assembly as a result. For the tails our recent discovery of the tail assembly chaperones and the way they interact with the tape measure protein and with the tail tube protein illuminates the main features of tail assembly in a way that was not previously clear. Both of our model systems also provide examples of how size can be determined in biological assembly. HK97 assembles T=7 capsids but D3, which has a very similar capsid protein sequence, assembles a T=9. The tail assembles to a length that we showed years ago is determined by the tape measure protein. Our recent result with the assembly chaperones clarifies how the length information in the tape measure protein is translated into tail length. Our proposed experiments with the HK97 heads will include generating mutants of the capsid protein chosen to alter the process of capsid maturation. These will be evaluated by their biochemical and structural phenotypes and interpreted in the context of the wealth of structural and structural/dynamic data available from earlier studies. We will pay particular attention to the Delta domain of the capsid protein, which has crucial roles in several aspects of capsid protein function. For tails we will concentrate initially on investigating the newly identified assembly intermediate, a complex of the tape measure protein and the two assembly chaperones, for which we will examine the structure and assembly activities. We will begin experiments to determine the structure and function of the tail tip, including the minor proteins and the newly discovered iron-sulfur cluster We propose to characterize two examples of non-canonical translation of a head protein (TerL), and to carry out a basic characterization of a newly discovered bacteriophage of a previously unknown type.

描述（由申请人提供）：本资助的主要重点是了解蛋白质纳米机器如何从其部件组装，组装后它如何运作，以及组装机制如何支持纳米机器的生物功能。为了解决这些问题，我们将使用噬菌体HK97的衣壳和噬菌体尾部？？这两种方法都为生物组装的核心重要方面提供了实验途径：HK97衣壳蛋白的Delta结构域是一种组装伴侣，它介导衣壳内衣壳体的组装，也结合蛋白酶并将其带入衣壳内部。它也有一个作用，使入口进入位置，并可能有一个关键的作用，在成核衣壳组装。对于尾部，我们最近发现的尾部装配伴侣及其与卷尺蛋白和尾管蛋白相互作用的方式，以一种以前不清楚的方式阐明了尾部装配的主要特征。我们的两个模型系统也提供了如何在生物组装中确定尺寸的例子。HK97组装T=7衣壳，而D3，具有非常相似的衣壳蛋白序列，组装T=9。尾巴的长度我们几年前就展示过是由卷尺蛋白决定的。我们最近对装配伴侣蛋白的研究结果阐明了卷尺蛋白中的长度信息是如何转化为尾巴长度的。我们提出的HK97头的实验将包括产生衣壳蛋白的突变体来改变衣壳成熟的过程。这些将通过其生化和结构表型进行评估，并在早期研究中提供的丰富的结构和结构/动态数据的背景下进行解释。我们将特别关注衣壳蛋白的Delta结构域，它在衣壳蛋白功能的几个方面起着至关重要的作用。对于尾部，我们将首先集中研究新发现的组装中间体，一个由卷尺蛋白和两个组装伴侣蛋白组成的复合体，我们将研究其结构和组装活性。我们将开始实验，以确定尾尖的结构和功能，包括次要蛋白质和新发现的铁硫簇。我们建议表征头部蛋白质（TerL）的两个非规范翻译的例子，并对新发现的一种以前未知类型的噬菌体进行基本表征。