Differential transcription profiling of the phage LUZ19 infection process in different growth media

RNA Biol. 2021 Nov;18(11):1778-1790. doi: 10.1080/15476286.2020.1870844. Epub 2021 Jan 15.

Abstract

RNA sequencing of phage-infected bacterial cultures offers a snapshot of transcriptional events occurring during the infection process, providing insights into the phage transcriptional organization as well as the bacterial response. To better mimic real environmental contexts, we performed RNA-seq of Pseudomonas aeruginosa PAO1 cultures infected with phage LUZ19 in a mammalian cell culture medium to better simulate a phage therapy event and the data were compared to lysogeny broth medium. Regardless of the media, phage LUZ19 induces significant transcriptional changes in the bacterial host over time, particularly during early infection (t = 5 min) and gradually shuts down bacterial transcription. In a common response in both media, 56 P. aeruginosa PAO1 genes are differentially transcribed and clustered into several functional categories such as metabolism, translation and transcription. Our data allowed us to tease apart a medium-specific response during infection from the identified infection-associated responses. This reinforces the concept that phages overtake bacterial transcriptome in a strict manner to gain control of the bacterial machinery and reallocate resources for infection, in this case overcoming the nutritional limitations of the mammalian cell culture medium. From a phage therapy perspective, this study contributes towards a better understanding of phage-host interaction in human physiological conditions and demonstrates the versatility of phage LUZ19 to adapt to different environments.

Keywords: Bacteriophage; P. aeruginosa; RNA-seq; gene expression; transcriptome.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Bacterial Proteins / genetics
  • Bacterial Proteins / metabolism*
  • Bacteriophages / physiology*
  • Cell Culture Techniques
  • Culture Media / pharmacology*
  • Gene Expression Regulation, Bacterial / drug effects*
  • Host-Pathogen Interactions*
  • Humans
  • Pseudomonas aeruginosa / drug effects
  • Pseudomonas aeruginosa / genetics*
  • Pseudomonas aeruginosa / growth & development
  • Pseudomonas aeruginosa / virology
  • Transcriptome*

Substances

  • Bacterial Proteins
  • Culture Media

Grants and funding

This study was supported by the Portuguese Foundation for Science and Technology (FCT) under the scope of the project PTDC/BBB-BSS/6471/2014 (POCI-01-0145-FEDER-016643) and the strategic funding of UIDB/04469/2020 unit. AB is supported by FCT through the grant SFRH/BD/133193/2017. This research was supported by funding from the European Research Council under the European Union’s Horizon 2020 research and innovation programme (Grant agreement No. 819800) awarded to RL.