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Viruses 2018, 10(6), 308; https://doi.org/10.3390/v10060308

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

1
Gene Expression and Regulation Section, Laboratory of Cell and Molecular Biology, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, Bethesda, MD 20814, USA
2
Section on Microbial Regulation, Eunice Kennedy Shriver National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, MD 20814, USA
3
Molecular Genomics Core, Eunice Kennedy Shriver National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, MD 20814, USA
*
Author to whom correspondence should be addressed.
Received: 28 March 2018 / Revised: 23 May 2018 / Accepted: 24 May 2018 / Published: 6 June 2018
(This article belongs to the Special Issue Phage-Host Interactions)
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Abstract

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. View Full-Text
Keywords: bacteriophage T4; DksA; MotA; RNA-seq; transcriptome analysis bacteriophage T4; DksA; MotA; RNA-seq; transcriptome analysis
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Patterson-West, J.; James, T.D.; Fernández-Coll, L.; Iben, J.R.; Moon, K.; Knipling, L.; Cashel, M.; Hinton, D.M. The E. coli Global Regulator DksA Reduces Transcription during T4 Infection. Viruses 2018, 10, 308.

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