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Summary   Pseudomonas  virus vB_PaeM_PA5oct is proposed as a model jumbo bacteriophage to investigate phage‐bacteria interactions and is a candidate for phage therapy applications. Combining hybrid sequencing, RNA‐Seq and mass spectrometry allowed us to accurately annotate its 286,783 bp genome with 461 coding regions including four non‐coding RNAs (ncRNAs) and 93 virion‐associated proteins. PA5oct relies on the host RNA polymerase for the infection cycle and RNA‐Seq revealed a gradual take‐over of the total cell transcriptome from 21% in early infection to 93% in late infection. PA5oct is not organized into strictly contiguous regions of temporal transcription, but some genomic regions transcribed in early, middle and late phases of infection can be discriminated. Interestingly, we observe regions showing limited transcription activity throughout the infection cycle. We show that PA5oct upregulates specific bacterial operons during infection including operons  pncA‐pncB1‐nadE  involved in NAD biosynthesis,  psl  for exopolysaccharide biosynthesis and  nap  for periplasmic nitrate reductase production. We also observe a downregulation of T4P gene products suggesting mechanisms of superinfection exclusion. We used the proteome of PA5oct to position our isolate amongst other phages using a gene‐sharing network. This integrative omics study illustrates the molecular diversity of jumbo viruses and raises new questions towards cellular regulation and phage‐encoded hijacking mechanisms.

Integrative omics analysis of Pseudomonas aeruginosa virus PA5oct highlights the molecular complexity of jumbo phages