Cryo-EM reveals infection steps of single-stranded RNA bacteriophages

Gorzelnik, Karl V.; Zhang, Junjie

doi:10.1016/j.pbiomolbio.2020.07.011

Cited by 7 publications

(3 citation statements)

References 69 publications

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“…There are 60 asymmetric A/B and 29 symmetric C/C CP 2 positions in its T = 3 capsid, with the additional C/ C lattice position occupied by MP. [40][41][42] The B conformation requires that an essential conserved Pro68 residue adopt a cis peptide bond, which is trans in A & C conformers. [43][44] PS-binding is distal from the side-chain of Pro68 21 and the allosteric effect of PS binding is propagated via its impact on the dynamics of the b-strands attached to the loop.…”

Section: Xrf Workflow and Choice Of Viral Footprinting Targetsmentioning

confidence: 99%

Genome-regulated Assembly of a ssRNA Virus May Also Prepare It for Infection

Chandler-Bostock

Bingham

Clark

et al. 2022

Journal of Molecular Biology

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Section: Xrf Workflow and Choice Of Viral Footprinting Targetsmentioning

confidence: 99%

Genome-regulated Assembly of a ssRNA Virus May Also Prepare It for Infection

Chandler-Bostock

Bingham

Clark

et al. 2022

Journal of Molecular Biology

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“…The icosahedral Qβ capsid enclosing the (+)-RNA genome is composed of 176 copies of the coat protein, 3-5 read-through proteins and one molecule of the maturation protein (10,11). During the rst step of the Qβ life cycle, the phage particle adsorbs to the surface of E. coli via contact between the maturation protein on the viral surface and the bacterial F-pilus (12,13). This interaction promotes the release of the Qβ (+)-RNA genome into the host cell via a poorly understood mechanism, which may rely on the retraction of the F-pilus to pull the complex of the maturation protein and the Qβ (+)-RNA into the host cell (13,14).…”

Section: Introductionmentioning

confidence: 99%

“…During the rst step of the Qβ life cycle, the phage particle adsorbs to the surface of E. coli via contact between the maturation protein on the viral surface and the bacterial F-pilus (12,13). This interaction promotes the release of the Qβ (+)-RNA genome into the host cell via a poorly understood mechanism, which may rely on the retraction of the F-pilus to pull the complex of the maturation protein and the Qβ (+)-RNA into the host cell (13,14). Next, the Qβ (+)-RNA is translated by host ribosomes in a tightly controlled process (15).…”

Section: Introductionmentioning

confidence: 99%

A Cell-free Infection System to Study Translation, Replication and Phage-particle Production During Infection of E. coli by Bacteriophage Qβ

Rasmussen

Knudsen

2021

Preprint

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Background Viruses infect all kingdoms of life, and new species are continuously being discovered. The single-stranded (+)-RNA viruses comprise the largest group of viruses, which includes pathogens such as Dengue virus, Corona virus and West Nile virus. Also, the simple bacteriophage Qβ belongs to this group of viruses. Studies of the mechanism of Qβ infection can increase our general understanding of the single-stranded (+)-RNA viruses, which can be exploited in the pursuit to treat and prevent of diseases caused by pathogenic (+)-RNA viruses.MethodsIn this study, we have analysed the production of infectious Qβ phage particles in three different cell-free infection systems upon addition of the Qβ genome as a template. The cell-free infection systems were based on cell-free protein expression systems: two commercial systems and one custom-made system. We studied the course of infection by analysing the production of viral RNA, proteins and phage particles produced in the cell-free reactions. The replication of the viral RNA was determined by RT-PCR, while the translation of the viral proteins was examined by radiolabelling, and the production of infectious phage particles was evaluated by double-layered plaque assays. ResultsBacteriophage Qβ was found to replicate in two of the three tested cell-free infection systems. Specifically, the viral RNA was replicated, the viral proteins were translated, and infectious phage particles were produced in the cell-free infection systems. The pattern of translation regulation of the viral proteins appeared similar to in vivo infection. Infectious Qβ phage particles were produced at yields of 25.4 × 104 PFU/μL reaction and 2.5 × 103 PFU/μL reaction in the commercial and custom-made system, respectively. Importantly, intact Qβ phage particles were shown not to replicate in the cell-free infection systems under the tested conditions. ConclusionCell-free infection systems can support replication of viral RNA, translation of viral proteins and self-assembly of infectious Qβ phage particles. We provide opportunities for further optimisation of the phage particle yield. Cell-free infection systems can be used in the future to study newly discovered viruses, the development of antiviral and antibacterial drugs, and in biotechnology.

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pH‐Responsive Virus‐Based Colloidal Crystals for Advanced Material Platforms

Tran,

Watts,

Valentin

et al. 2024

Adv Funct Materials

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Bacteriophages have a well‐defined nanoscale size, shape, and surface chemistry, making them promising candidates for creating advanced biomaterials for applications including biocatalysis, drug delivery, and biosensing. This study demonstrates the self‐assembly of the ≈29 nm diameter bacteriophage Qbeta (Qubevirus durum) with the synthetic polycation, poly [2‐(methacryloyloxy)ethyl] trimethylammonium chloride (pMETAC), into compartmentalized colloidal crystals. The pH and the polymer chain length tune their self‐assembly and the resulting structure, with the potential for further chemical modification or loading with bioactive molecules. Small angle X‐ray scattering (SAXS), multi‐angle dynamic light scattering (DLS), and atomic force microscopy (AFM) are used for studying the Qbeta self‐assembly into the geometrically ordered aggregates. The suprastructures form at pH > 7.0 and disassemble at pH < 7.0. Zeta potential measurements and X‐ray photoelectron spectroscopy (XPS) show pMETAC adsorption onto the negatively charged Qbeta surface. The colloidal crystal formation is achieved without chemically modifying the Qbeta surface. Additionally, the Qbeta/pMETAC suprastructures can be easily separated from the suspension as macroscopic aggregate, maintaining their activity. Their simple preparation allows for large‐scale production of advanced materials in food and health science applications and nanotechnology. The insights from this study will further advance the tailored design and production of novel colloidal materials.

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Cryo-EM reveals infection steps of single-stranded RNA bacteriophages

Cited by 7 publications

References 69 publications

Genome-regulated Assembly of a ssRNA Virus May Also Prepare It for Infection

Genome-regulated Assembly of a ssRNA Virus May Also Prepare It for Infection

A Cell-free Infection System to Study Translation, Replication and Phage-particle Production During Infection of E. coli by Bacteriophage Qβ

pH‐Responsive Virus‐Based Colloidal Crystals for Advanced Material Platforms

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