Sequential membrane- and protein-bound organelles compartmentalize genomes during phage infection
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- Evaluated articles (Rapid Reviews Infectious Diseases)
- Evaluated articles (Arcadia Science)
Abstract
Eukaryotic viruses assemble compartments required for genome replication, but no such organelles are known to be essential for prokaryotic viruses. Bacteriophages of the family Chimalliviridae sequester their genomes within a phagegenerated organelle, the phage nucleus, which is enclosed by a lattice of viral protein ChmA. Using the dRfxCas13d-based knockdown system CRISPRi-ART, we show that ChmA is essential for the E. coli phage Goslar life cycle. Without ChmA, infections are arrested at an early stage in which the injected phage genome is enclosed in a membrane-bound vesicle capable of gene expression but not DNA replication. Not only do we demonstrate that the phage nucleus is essential for genome replication, but we also show that the Chimalliviridae early phage infection (EPI) vesicle is a transcriptionally active, phage-generated organelle.
Article activity feed
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Kevin Forsberg
Review 3: "Sequential Membrane- and Protein-bound Organelles Compartmentalize Genomes During Phage Infection"
Overall, the reviews provide critical scrutiny of the methodology and balanced perspective on the merits and limitations of this preprint's conclusions.
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Ekaterina Semenova, Hiba Shaqra
Review 2: "Sequential Membrane- and Protein-bound Organelles Compartmentalize Genomes During Phage Infection"
Overall, the reviews provide critical scrutiny of the methodology and balanced perspective on the merits and limitations of this preprint's conclusions.
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Maria Yakunina, Daria Antonova
Review 1: "Sequential Membrane- and Protein-bound Organelles Compartmentalize Genomes During Phage Infection"
Overall, the reviews provide critical scrutiny of the methodology and balanced perspective on the merits and limitations of this preprint's conclusions.
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Strength of evidence
Reviewers: M Yakunina & D Antonova (Peter the Great St.Petersburg Polytechnic University) | 📗📗📗📗◻️
E Semenova & H Shaqra (Rutgers University) | 📘📘📘📘📘
K Forsberg (University of Texas) | 📘📘📘📘📘 -
viral mRNAs are exported to the cytoplasm to be translated by host ribosomes
Based on Figure 1B, it seems like you are suggesting that knockdown occurs after the phage nucleus already forms, but this seems contradictory to rest of the results. So maybe 1B is more of a generic overview of how this knockdown technology is particularly useful in nucleus-forming phage. However, since in this paper the knockdown is presumably happening before nucleus formation, it might be good to have a figure somewhere reflecting that, as I found 1B to be a little confusing in the context of ChmA.
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reduced ∼1,000-fold
I'm curious what the thresholds are for considering something to be essential for phage reproduction? 1000x reduction is huge, but there is still a decent amount of plaquing shown in figure 1F.
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the EPI vesicle is transcriptionally active and is a bona fide stage in the Chimalliviridae life cycle
this is so interesting! I wonder whether the EPI vesicle is found in non-nucleus forming jumbo phage? Is it maybe an intermediate step in the evolution of these phage?
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Thus, Chimalliviridae form sophisticated subcellular compartments of distinct compositions and functions that facilitate successive stages of the life cycle.
Congrats on a really beautiful paper! So much cool new biology!
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