Introducing differential RNA-seq mapping to track the early infection phase for Pseudomonas phage ɸKZ

RNA Biol. 2021 Aug;18(8):1099-1110. doi: 10.1080/15476286.2020.1827785. Epub 2020 Oct 25.

Abstract

As part of the ongoing renaissance of phage biology, more phage genomes are becoming available through DNA sequencing. However, our understanding of the transcriptome architecture that allows these genomes to be expressed during host infection is generally poor. Transcription start sites (TSSs) and operons have been mapped for very few phages, and an annotated global RNA map of a phage - alone or together with its infected host - is not available at all. Here, we applied differential RNA-seq (dRNA-seq) to study the early, host takeover phase of infection by assessing the transcriptome structure of Pseudomonas aeruginosa jumbo phage ɸKZ, a model phage for viral genetics and structural research. This map substantially expands the number of early expressed viral genes, defining TSSs that are active ten minutes after ɸKZ infection. Simultaneously, we record gene expression changes in the host transcriptome during this critical metabolism conversion. In addition to previously reported upregulation of genes associated with amino acid metabolism, we observe strong activation of genes with functions in biofilm formation (cdrAB) and iron storage (bfrB), as well as an activation of the antitoxin ParD. Conversely, ɸKZ infection rapidly down-regulates complexes IV and V of oxidative phosphorylation (atpCDGHF and cyoABCDE). Taken together, our data provide new insights into the transcriptional organization and infection process of the giant bacteriophage ɸKZ and adds a framework for the genome-wide transcriptomic analysis of phage-host interactions.

Keywords: Bacteriophage ɸKZ; Pseudomonas aeruginosa; dRNA-seq; differential expression; phage-host interaction; transcription start site.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adhesins, Bacterial / genetics
  • Adhesins, Bacterial / metabolism
  • Antibiosis / genetics*
  • Bacterial Outer Membrane Proteins / genetics
  • Bacterial Outer Membrane Proteins / metabolism
  • Bacterial Proteins / genetics
  • Bacterial Proteins / metabolism
  • Biofilms / growth & development
  • Chromosome Mapping
  • DNA-Binding Proteins / genetics
  • DNA-Binding Proteins / metabolism
  • Gene Expression Regulation, Bacterial*
  • Gene Expression Regulation, Viral*
  • Gene Ontology
  • Genome, Viral*
  • Molecular Sequence Annotation
  • Pseudomonas Phages / genetics*
  • Pseudomonas Phages / growth & development
  • Pseudomonas Phages / metabolism
  • Pseudomonas aeruginosa / genetics*
  • Pseudomonas aeruginosa / metabolism
  • Pseudomonas aeruginosa / virology
  • RNA, Viral / genetics
  • RNA, Viral / metabolism
  • Sequence Analysis, RNA
  • Transcription Initiation Site
  • Transcriptome

Substances

  • Adhesins, Bacterial
  • Bacterial Outer Membrane Proteins
  • Bacterial Proteins
  • CdrA protein, Pseudomonas aeruginosa
  • DNA-Binding Proteins
  • RNA, Viral
  • parD protein, Bacteria

Grants and funding

LW holds a predoctoral scholarship from FWO-fundamental research (11D8920N). We thank the Vogel Stiftung Dr. Eckernkamp for supporting FP with a Dr. Eckernkamp Fellowship. This article is part of a project that has received funding from the European Research Council (ERC) under the European Union’s ERC consolidator grant awarded to RL (Grant agreement No. [819800])