Antibacterial phage ORFans of Pseudomonas aeruginosa phage LUZ24 reveal a novel MvaT inhibiting protein

Wagemans, Jeroen; Delattre, Anne‐Sophie; Uytterhoeven, Birgit; Smet, Jeroen De; Cenens, William; Aertsen, Abram; Ceyssens, Pieter‐Jan; Lavigne, Rob

doi:10.3389/fmicb.2015.01242

Cited by 37 publications

(45 citation statements)

References 52 publications

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“…In addition, bacterial histone-like proteins, such as H-NS in E. coli , can repress viral transcription (reviewed in [ 79 ]). Interestingly, the T7 protein gp5.5 [ 80 ], the T4 protein Arn [ 81 ], and the phage LUZ24 protein MvaT [ 82 ] have each been shown to inhibit H-NS repression through distinct mechanisms. Future work investigating the effects of bacterial transcriptional regulators will likely reveal additional host proteins that regulate viral gene expression.…”

Section: Discussionmentioning

confidence: 99%

The E. coli Global Regulator DksA Reduces Transcription during T4 Infection

Patterson-West

James

Fernández-Coll

et al. 2018

Viruses

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Bacteriophage T4 relies on host RNA polymerase to transcribe three promoter classes: early (Pe, requires no viral factors), middle (Pm, requires early proteins MotA and AsiA), and late (Pl, requires middle proteins gp55, gp33, and gp45). Using primer extension, RNA-seq, RT-qPCR, single bursts, and a semi-automated method to document plaque size, we investigated how deletion of DksA or ppGpp, two E. coli global transcription regulators, affects T4 infection. Both ppGpp0 and ΔdksA increase T4 wild type (wt) plaque size. However, ppGpp0 does not significantly alter burst size or latent period, and only modestly affects T4 transcript abundance, while ΔdksA increases burst size (2-fold) without affecting latent period and increases the levels of several Pe transcripts at 5 min post-infection. In a T4motAam infection, ΔdksA increases plaque size and shortens latent period, and the levels of specific middle RNAs increase due to more transcription from Pe’s that extend into these middle genes. We conclude that DksA lowers T4 early gene expression. Consequently, ΔdksA results in a more productive wt infection and ameliorates the poor expression of middle genes in a T4motAam infection. As DksA does not inhibit Pe transcription in vitro, regulation may be indirect or perhaps requires additional factors.

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Section: Discussionmentioning

confidence: 99%

The E. coli Global Regulator DksA Reduces Transcription during T4 Infection

Patterson-West

James

Fernández-Coll

et al. 2018

Viruses

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“…However, Arn is not essential [ 11 ] and whether its absence affects T4 gene expression is not known. Specific H-NS antagonists have also been identified for T7 [ 12 ], Luz24 [ 13 ], and Mu [ 14 , 15 ].…”

Section: Introductionmentioning

confidence: 99%

The Bacteriophage T4 MotB Protein, a DNA-Binding Protein, Improves Phage Fitness

Patterson-West

Arroyo-Mendoza

Hsieh

et al. 2018

Viruses

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The lytic bacteriophage T4 employs multiple phage-encoded early proteins to takeover the Escherichia coli host. However, the functions of many of these proteins are not known. In this study, we have characterized the T4 early gene motB, located in a dispensable region of the T4 genome. We show that heterologous production of MotB is highly toxic to E. coli, resulting in cell death or growth arrest depending on the strain and that the presence of motB increases T4 burst size 2-fold. Previous work suggested that motB affects middle gene expression, but our transcriptome analyses of T4 motBam vs. T4 wt infections reveal that only a few late genes are mildly impaired at 5 min post-infection, and expression of early and middle genes is unaffected. We find that MotB is a DNA-binding protein that binds both unmodified host and T4 modified [(glucosylated, hydroxymethylated-5 cytosine, (GHme-C)] DNA with no detectable sequence specificity. Interestingly, MotB copurifies with the host histone-like proteins, H-NS and StpA, either directly or through cobinding to DNA. We show that H-NS also binds modified T4 DNA and speculate that MotB may alter how H-NS interacts with T4 DNA, host DNA, or both, thereby improving the growth of the phage.

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“…Each fragment was tested both as bait and prey protein, thus raising the reliability of the screen. To verify the efficiency of this pooled array approach, a positive control was included in the screen, using interacting proteins Luz24_gp4 (pGBT9) and MvaT (pGAD424) (Wagemans et al, 2015). …”

Section: Resultsmentioning

confidence: 99%

A Protein Interaction Map of the Kalimantacin Biosynthesis Assembly Line

et al. 2016

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The antimicrobial secondary metabolite kalimantacin (also called batumin) is produced by a hybrid polyketide/non-ribosomal peptide system in Pseudomonas fluorescens BCCM_ID9359. In this study, the kalimantacin biosynthesis gene cluster is analyzed by yeast two-hybrid analysis, creating a protein–protein interaction map of the entire assembly line. In total, 28 potential interactions were identified, of which 13 could be confirmed further. These interactions include the dimerization of ketosynthase domains, a link between assembly line modules 9 and 10, and a specific interaction between the trans-acting enoyl reductase BatK and the carrier proteins of modules 8 and 10. These interactions reveal fundamental insight into the biosynthesis of secondary metabolites. This study is the first to reveal interactions in a complete biosynthetic pathway. Similar future studies could build a strong basis for engineering strategies in such clusters.

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Antibacterial phage ORFans of Pseudomonas aeruginosa phage LUZ24 reveal a novel MvaT inhibiting protein

Cited by 37 publications

References 52 publications

The E. coli Global Regulator DksA Reduces Transcription during T4 Infection

The E. coli Global Regulator DksA Reduces Transcription during T4 Infection

The Bacteriophage T4 MotB Protein, a DNA-Binding Protein, Improves Phage Fitness

A Protein Interaction Map of the Kalimantacin Biosynthesis Assembly Line

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