Phage T5 two-step injection

Davison, John

doi:10.1101/866236

Cited by 3 publications

(1 citation statement)

References 42 publications

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“…The majority of the suppressor mutations identified were missense mutations, although some included frameshifts, nonsense, and promoter mutations ( Tables S4 and S5 ). Genes orf008 and orf015 are encoded in the “first-step transfer” region of the T5 genome, an 11 kb section that is injected first into host bacteria and coordinates injection pausing for approximately five minutes before the remainder of the genome is injected (Davison, 2019). During those first minutes of infection, other pre-early genes of the first-step transfer region remodel core processes and shut down signaling within the host cell (Davison, 2015).…”

Section: Resultsmentioning

confidence: 99%

Bacterial NLR-related proteins protect against phage

Kibby

Conte

Burroughs

et al. 2022

Preprint

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Bacteria use a wide range of immune systems to counter phage infection. A subset of these genes share homology with components of eukaryotic immune systems, suggesting that eukaryotes horizontally acquired certain innate immune genes from bacteria. Here we show that proteins containing a NACHT module, the central feature of the animal nucleotide-binding domain and leucine-rich repeat containing gene family (NLRs), are found in bacteria and defend against phages. NACHT proteins are widespread in bacteria, provide immunity against both DNA and RNA phages, and display the characteristic C-terminal sensor, central NACHT, and N-terminal effector modules. Some bacterial NACHT proteins have domain architectures similar to human NLRs that are critical components of inflammasomes. Human disease-associated NLR mutations that cause stimulus-independent activation of the inflammasome also activate bacterial NACHT proteins, supporting a shared signaling mechanism. This work establishes that NACHT module-containing proteins are ancient mediators of innate immunity across the tree of life.

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Section: Resultsmentioning

confidence: 99%

Bacterial NLR-related proteins protect against phage

Kibby

Conte

Burroughs

et al. 2022

Preprint

View full text Add to dashboard Cite

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Phage T5 two-step injection

Davison

2019

Preprint

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Escherichia coli bacteriophage T5 differs from most phages in that it injects its genome in two steps: First Step Transfer, FST, corresponding to leftmost 7.9% of the genome and Second Step Transfer, SST, corresponding to the remainder of the genome. Expression of genes A1 and A2 is required for SST. DNA injection stops at a site known as the injection stop signal (iss) which is a cis acting site located in the untranslated region of the Left Terminal Repeat (LTR). The iss region is extremely complicated with many repeats, inverted repeats and palindromes. This report compares the iss regions of 21 T5 related phages and shows that all have a common conserved structure including a series of 8 DnaA boxes arranged in a highly specific manner; reminiscent of the origin of replication (oriC) of the host. DnaA protein, which binds DnaA boxes is a mostly membrane bound, leading to the suggestion that injection stops at iss due to binding to DnaA protein at the membrane. A new model of the mechanism of T5 injection stop and SST injection re-start is suggested.

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Editing of Phage Genomes—Recombineering-assisted SpCas9 Modification of Model Coliphages T7, T5, and T3

et al. 2022

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Bacteriophages—viruses that infect bacterial cells—are the most abundant biological entities on Earth. The use of phages in fundamental research and industry requires tools for precise manipulation of their genomes. Yet, compared to bacterial genome engineering, modification of phage genomes is challenging because of the lack of selective markers and thus requires laborious screenings of recombinant/mutated phage variants. The development of the CRISPR-Cas technologies allowed to solve this issue by the implementation of negative selection that eliminates the parental phage genomes. In this manuscript, we summarize current methods of phage genome engineering and their coupling with CRISPR-Cas technologies. We also provide examples of our successful application of these methods for introduction of specific insertions, deletions, and point mutations in the genomes of model Escherichia coli lytic phages T7, T5, and T3.

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Phage T5 two-step injection

Cited by 3 publications

References 42 publications

Bacterial NLR-related proteins protect against phage

Bacterial NLR-related proteins protect against phage

Phage T5 two-step injection

Editing of Phage Genomes—Recombineering-assisted SpCas9 Modification of Model Coliphages T7, T5, and T3

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