A Phage Tubulin Assembles Dynamic Filaments by an Atypical Mechanism to Center Viral DNA within the Host Cell

Kraemer, James A.; Erb, Marcella L.; Waddling, C.A.; Montabana, Elizabeth; Zehr, Elena A.; Wang, Hannah; Nguyen, Katrina; Pham, Duy Stephen L.; Agard, David A.; Pogliano, Joe

doi:10.1016/j.cell.2012.04.034

Cited by 102 publications

(253 citation statements)

References 44 publications

(58 reference statements)

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“…Unfortunately, the tails were not resolved in recent EM TubZ filament reconstructions (16), but it can be deduced from the structures that the filaments will most likely have TubR-binding tails exposed at one end only. The accessibility of TubZ tails for TubRC centromeric complexes at one end of the filament assumes that the tails also bind along the filament, latching onto subunits further up as has clearly been seen in phage TubZ (PhuZ) protofilament crystal structures (26,27). In accordance with competitive TubZ tail binding between the filament lattice and TubRC, we have observed lateral interaction of TubRC with filaments.…”

Section: Discussionsupporting

confidence: 65%

Reconstitution of a prokaryotic minus end-tracking system using TubRC centromeric complexes and tubulin-like protein TubZ filaments

Fink

Löwe

2015

Proc. Natl. Acad. Sci. U.S.A.

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Segregation of DNA is a fundamental process during cell division. The mechanism of prokaryotic DNA segregation is largely unknown, but several low-copy-number plasmids encode cytomotive filament systems of the actin type and tubulin type important for plasmid inheritance. Of these cytomotive filaments, only actin-like systems are mechanistically well characterized. In contrast, the mechanism by which filaments of tubulin-like TubZ protein mediate DNA motility is unknown. To understand polymer-driven DNA transport, we reconstituted the filaments of TubZ protein (TubZ filaments) from Bacillus thuringiensis pBtoxis plasmid with their centromeric TubRC complexes containing adaptor protein TubR and tubC DNA. TubZ alone assembled into polar filaments, which annealed laterally and treadmilled. Using single-molecule imaging, we show that TubRC complexes were not pushed by filament polymerization; instead, they processively tracked shrinking, depolymerizing minus ends. Additionally, the TubRC complex nucleated TubZ filaments and allowed for treadmilling. Overall, our results indicate a pulling mechanism for DNA transport by the TubZRC system. The discovered minus end-tracking property of the TubRC complex expands the mechanistic diversity of the prokaryotic cytoskeleton.cytoskeleton | tubulin homologue | DNA segregation

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Section: Discussionsupporting

confidence: 65%

Reconstitution of a prokaryotic minus end-tracking system using TubRC centromeric complexes and tubulin-like protein TubZ filaments

Fink

Löwe

2015

Proc. Natl. Acad. Sci. U.S.A.

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“…This is further illustrated by the enrichment of peptidoglycan and lipopolysaccharide biosynthesis during PEV2, LUZ24 and phiKZ infection. In fact, Phikzlikevirus members encode a tubulin-like protein (Kraemer et al, 2012), which positions the phage DNA in the center of the cell and induces an arrest in host cell division during infection . Other cell division inhibiting early proteins were also identified in LUZ7, an N4likevirus like PEV2 (Wagemans et al, 2014).…”

Section: Resultsmentioning

confidence: 99%

High coverage metabolomics analysis reveals phage-specific alterations to Pseudomonas aeruginosa physiology during infection

et al. 2016

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Phage-mediated metabolic changes in bacteria are hypothesized to markedly alter global nutrient and biogeochemical cycles. Despite their theoretic importance, experimental data on the net metabolic impact of phage infection on the bacterial metabolism remains scarce. In this study, we tracked the dynamics of intracellular metabolites using untargeted high coverage metabolomics in Pseudomonas aeruginosa cells infected with lytic bacteriophages from six distinct phage genera. Analysis of the metabolomics data indicates an active interference in the host metabolism. In general, phages elicit an increase in pyrimidine and nucleotide sugar metabolism. Furthermore, clear phage-specific and infection stage-specific responses are observed, ranging from extreme metabolite depletion (for example, phage YuA) to complete reorganization of the metabolism (for example, phage phiKZ). As expected, pathways targeted by the phage-encoded auxiliary metabolic genes (AMGs) were enriched among the metabolites changing during infection. The effect on pyrimidine metabolism of phages encoding AMGs capable of host genome degradation (for example, YuA and LUZ19) was distinct from those lacking nuclease-encoding genes (for example, phiKZ), which demonstrates the link between the encoded set of AMGs of a phage and its impact on host physiology. However, a large fraction of the profound effect on host metabolism could not be attributed to the phage-encoded AMGs. We suggest a potentially crucial role for small, 'non-enzymatic' peptides in metabolism take-over and hypothesize on potential biotechnical applications for such peptides. The highly phage-specific nature of the metabolic impact emphasizes the potential importance of the 'phage diversity' parameter when studying metabolic interactions in complex communities.

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“…29 Another "large" phage known to be in this category is Pseudomonas chlororaphis phage 201phi2-1, a lytic phage in the phiKZ family, genome length D 316.674 Kb. [34][35][36] The largest known phage, G (GenBank: genome length D 497.513 Kb), is similar. Phage G propagates poorly in liquid culture and very well in medium with a dilute agarose gel.…”

Section: Improving the Propagation Of 0305phi8-36 And Other Large Phagesmentioning

confidence: 91%

Enhancing and initiating phage-based therapies

et al. 2014

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D rug development has typically been a primary foundation of strategy for systematic, long-range management of pathogenic cells. However, drug development is limited in speed and flexibility when response is needed to changes in pathogenic cells, especially changes that produce drug-resistance. The high replication speed and high diversity of phages are potentially useful for increasing both response speed and response flexibility when changes occur in either drug resistance or other aspects of pathogenic cells. We present strategy, with some empirical details, for (1) using modern molecular biology and biophysics to access these advantages during the phage therapy of bacterial infections, and (2) initiating use of phage capsid-based drug delivery vehicles (DDVs) with procedures that potentially overcome both drug resistance and other present limitations in the use of DDVs for the therapy of neoplasms. The discussion of phage therapy includes (a) historical considerations, (b) changes that appear to be needed in clinical tests if use of phage therapy is to be expanded, (c) recent work on novel phages and its potential use for expanding the capabilities of phage therapy and (d) an outline for a strategy that encompasses both theory and practice for expanding the applications of phage therapy. The discussion of DDVs starts by reviewing current work on DDVs, including work on both liposomal and viral DDVs. The discussion concludes with some details of the potential use of permeability constrained phage capsids as DDVs.

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A Phage Tubulin Assembles Dynamic Filaments by an Atypical Mechanism to Center Viral DNA within the Host Cell

Cited by 102 publications

References 44 publications

Reconstitution of a prokaryotic minus end-tracking system using TubRC centromeric complexes and tubulin-like protein TubZ filaments

Reconstitution of a prokaryotic minus end-tracking system using TubRC centromeric complexes and tubulin-like protein TubZ filaments

High coverage metabolomics analysis reveals phage-specific alterations to Pseudomonas aeruginosa physiology during infection

Enhancing and initiating phage-based therapies

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