Journal Article

Cryo-EM analysis of Pseudomonas phage Pa193 structural components

Lemire, Sebastien, Iglesias, Stephano M., Sim, Lucy, Whitelegge, Julian, Birx, Deborah, Schauer, Evan, Kyme, Pierre, Cingolani, Gino, Reid, Johnny, Gao, Lucy, Hou, Chun-Feng David, Soriaga, Angela, Geier, Renae

2024

Overview

The World Health Organization has designated Pseudomonas aeruginosa as a critical pathogen for the development of new antimicrobials. Bacterial viruses, or bacteriophages, have been used in various clinical settings, commonly called phage therapy, to address this growing public health crisis. Here, we describe a high-resolution structural atlas of a therapeutic, contractile-tailed Pseudomonas phage, Pa193. We used bioinformatics, proteomics, and cryogenic electron microscopy single particle analysis to identify, annotate, and build atomic models for 21 distinct structural polypeptide chains forming the icosahedral capsid, neck, contractile tail, and baseplate. We identified a putative scaffolding protein stabilizing the interior of the capsid 5-fold vertex. We also visualized a large portion of Pa193 ~ 500 Å long tail fibers and resolved the interface between the baseplate and tail fibers. The work presented here provides a framework to support a better understanding of phages as biomedicines for phage therapy and inform engineering opportunities. Leveraging bioinformatics, proteomics, and cryogenic electron microscopy, this study deciphers the architecture and design principles of the therapeutic Pseudomonas phage Pa193.

• Publisher: Nature Publishing Group UK,Nature Publishing Group,Nature Portfolio
• Subject: 101/28 / 101/58 / 631/45/535/1258/1259 / 631/45/612/1256 / Antimicrobial agents / Bioinformatics / Biomedical and Life Sciences / Capsid Proteins - chemistry / Capsid Proteins - metabolism / Capsid Proteins - ultrastructure / Contractility / Cryoelectron Microscopy - methods / Electron microscopy / Fibers / Life Sciences / Microscopy / Models, Molecular / Phages / Protein folding / Proteomics / Pseudomonas / Pseudomonas aeruginosa - virology / Pseudomonas Phages - ultrastructure / Public health / Tail fiber protein