Molecular and cellular biology of the phage nucleus and spindle

噬菌体核和纺锤体的分子和细胞生物学

• 批准号: 10710178
• 负责人: JOSEPH A POGLIANO
• 金额: $ 49.32万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN DIEGO
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-09-06 至 2026-07-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10710178
• 关键词: Bacteriophages; Biochemistry; Biology; Capsid; Cell Nucleus; Cells; Cellular biology; Complex; Cryo-electron tomography; Cytoplasm; DNA Packaging; DNA Repair; DNA biosynthesis; Data; Docking; Escherichia coli; Eukaryota; Filament; Genetic; Genetic Recombination; Genetic Transcription; Genome; Goals; In Situ; In Vitro; Individual; Infection; Knowledge; Membrane; Membrane Lipids; Messenger RNA; Metabolism; Molecular and Cellular Biology; Play; Process; Protein Biosynthesis; Protein Import; Proteins; Pseudomonas; Pseudomonas aeruginosa; Research; Resolution; Role; Structure; Surface; Translation Process; Tubulin; Viral; Virus; Work; biological systems; candidate identification; design; insight; knock-down; live cell imaging; mRNA Export; nucleotide metabolism; protein expression; protein metabolism; segregation; self assembly; structural biology

Project Summary The ability to establish and maintain subcellular organization is a fundamental design principle of biological systems. We recently discovered that many jumbo phage establish surprisingly complex subcellular organization involving a nucleus-like structure or “phage nucleus” that compartmentalizes phage replication within the host cytoplasm. The phage nucleus carries out many functions previously only attributed to the eukaryotic nucleus including segregation of key processes (DNA replication and repair, mRNA synthesis, protein synthesis and metabolism) and selective protein import and mRNA export. However, the phage nucleus is structurally dissimilar to the eukaryotic nucleus, being mainly composed of a single layer of a phage-encoded protein and lacking evident openings for exchange with the cytoplasm instead of a double-layer lipid membrane containing prominent pore complexes. We have identified the key protein that makes up the phage nucleus shell; however, the components required to assemble a fully functional phage nucleus and their roles in this multipurpose structure remain unknown. Here we aim to identify and characterize additional proteins that are required to construct a fully functional phage nucleus to gain greater insight into the biology of these understudied viruses and the underlying principles of subcellular compartmentalization previously observed almost exclusively in eukaryotes. We will use an integrative approach that combines genetics, cell biology, biochemistry and in situ and in vitro structural biology approaches to study the jumbo phage in Pseudomonas and E. coli. Together, the knowledge gained from this project will provide insight into fundamental principles underlying subcellular organization.

项目概要 建立和维持亚细胞组织的能力是生物的基本设计原则 系统。我们最近发现许多巨型噬菌体建立了极其复杂的亚细胞组织 涉及核样结构或“噬菌体核”，将噬菌体在宿主内复制区分开来 细胞质。噬菌体核执行许多以前仅归属于真核细胞核的功能 包括关键过程的分离（DNA 复制和修复、mRNA 合成、蛋白质合成和 代谢）和选择性蛋白质输入和 mRNA 输出。然而，噬菌体核在结构上不同 对于真核细胞核来说，主要由单层噬菌体编码的蛋白质组成，缺乏 与细胞质交换的明显开口，而不是含有突出的双层脂质膜 孔隙复合体。我们已经确定了构成噬菌体核壳的关键蛋白质；然而， 组装功能齐全的噬菌体核所需的组件及其在这种多用途结构中的作用 仍然未知。在这里，我们的目标是识别和表征构建 功能齐全的噬菌体核，以更深入地了解这些正在研究的病毒的生物学和 以前几乎仅在真核生物中观察到的亚细胞区室化的基本原理。 我们将采用结合遗传学、细胞生物学、生物化学以及原位和体外的综合方法 研究假单胞菌和大肠杆菌中巨型噬菌体的结构生物学方法。在一起，知识 从这个项目中获得的知识将深入了解亚细胞组织的基本原理。

项目成果

A phage nucleus-associated RNA-binding protein is required for jumbo phage infection.

巨噬菌体感染需要噬菌体核相关 RNA 结合蛋白。

• DOI: 10.1101/2023.09.22.559000
• 发表时间: 2023
• 期刊: bioRxiv : the preprint server for biology
• 作者: Enustun,Eray;Armbruster,EmilyG;Lee,Jina;Zhang,Sitao;Yee,BrianA;Gu,Yajie;Deep,Amar;Naritomi,JackT;Liang,Qishan;Aigner,Stefan;Adler,BenjaminA;Cress,BradyF;Doudna,JenniferA;Chaikeeratisak,Vorrapon;Cleveland,DonW;Ghassemia
• 通讯作者: Ghassemia

Selective transport of fluorescent proteins into the phage nucleus.

• DOI: 10.1371/journal.pone.0251429
• 发表时间: 2021
• 期刊: PloS one
• 影响因子: 3.7
• 作者: Nguyen KT;Sugie J;Khanna K;Egan ME;Birkholz EA;Lee J;Beierschmitt C;Villa E;Pogliano J
• 通讯作者: Pogliano J