The Xp10 Bacteriophage Protein P7 Inhibits Transcription by the Major and Major Variant Forms of the Host RNA Polymerase via a Common Mechanism

• Published in: Journal of molecular biology Vol. 428; no. 20; pp. 3911 - 3919
• Main Authors: Brown, D.R., Sheppard, C.M., Burchell, L., Matthews, S., Wigneshweraraj, S.
• Format: Journal Article
• Language: English
• Published: England Elsevier Ltd 09.10.2016; Elsevier
• Subjects: bacteria; Bacterial Proteins - antagonists & inhibitors; bacteriophage; Bacteriophages - growth & development; Communication; DNA-Directed RNA Polymerases - antagonists & inhibitors; Host-Parasite Interactions; Promoter Regions, Genetic; Protein Binding; RNA polymerase; Sigma Factor - metabolism; transcription regulation; Transcription, Genetic; Viral Proteins - metabolism; Xanthomonas - genetics; Xanthomonas - virology; σ factor; RPo; transcription regulation; RPc; CBD; bacteria; β′ NTD; RNA polymerase; σ factor; RNAP; bacteriophage
• ISSN: 0022-2836; 1089-8638
• DOI: 10.1016/j.jmb.2016.08.004

The σ factor is a functionally obligatory subunit of the bacterial transcription machinery, the RNA polymerase. Bacteriophage-encoded small proteins that either modulate or inhibit the bacterial RNAP to allow the temporal regulation of bacteriophage gene expression often target the activity of the major bacterial σ factor, σ70. Previously, we showed that during Xanthomonas oryzae phage Xp10 infection, the phage protein P7 inhibits the host RNAP by preventing the productive engagement with the promoter and simultaneously displaces the σ70 factor from the RNAP. In this study, we demonstrate that P7 also inhibits the productive engagement of the bacterial RNAP containing the major variant bacterial σ factor, σ54, with its cognate promoter. The results suggest for the first time that the major variant form of the host RNAP can also be targeted by bacteriophage-encoded transcription regulatory proteins. Since the major and major variant σ factor interacting surfaces in the RNAP substantially overlap, but different regions of σ70 and σ54 are used for binding to the RNAP, our results further underscore the importance of the σ–RNAP interface in bacterial RNAP function and regulation and potentially for intervention by antibacterials. Graphical abstract [Display omitted] •Xp10 phage transcription regulator P7 inhibits transcription by RNAP containing σ54.•P7 prevents the productive engagement of the σ54–RNAP with the promoter DNA.• P7 disrupts preformed σ54–RNAP-promoter complexes.