Poster Presentation 23rd Annual Lorne Proteomics Symposium 2018

Temporal quantitative profiling of ΦX174 and host-cell proteomes during lytic infection (#127)

Bradley W Wright 1 , Dominic Y Logel 1 , Dana Pascovici 2 , Mehdi Mirzaei 1 2 , Paul R Jaschke 1 , Mark P Molloy 1 2
  1. Molecular Sciences, Macquarie University, Sydney, NSW, Australia
  2. Australian Proteome Analysis Facility, Macquarie University, NSW, Australia

The potential for bacteriophage to be used for industrial and medical purposes are on the cusp of breaking onto the market and to be able to model, control, engineer, and use viruses in medical and industrial purposes we must be able to measure their phenotypic responses over time. Unfortunately, there is currently limited understanding regarding host-cell proteomic response to lytic bacteriophage infection. This study explores lytic bacteriophage infection using the simple 11-gene model bacteriophage ΦX174 and its host Escherichia coli (strain C). A 10-plex TMT quantitative LC-MS strategy was used to probe the proteomic response to the ΦX174 infection across a five-point time-course covering the entirety of the 75-minute lytic infection.

2185 non-redundant proteins were identified at 1% false-discovery rate (FDR), 1764 of which were quantifiable across all samples. Using two-sample t-tests with multiple testing corrections, 706 proteins were observed to be differentially regulated (q-value < 0.05) across grouped time-points (early-stage, mid-stage, and late-stage) and/or phage infected and control samples. All 11 ΦX174 proteins were quantifiably detected, including the previously elusive phage lysis protein E. This enabled temporal mapping of ΦX174 proteome abundance during infection. Interestingly, we observed two host E. coli heat shock proteins, IbpA and IbpB, that are the dominant response to bacteriophage expression and have not previously been reported in this context. We are now undertaking functional categorization to reveal the host-cell response to infection.