Poster Presentation Lorne Infection and Immunity 2019

The role of the RNA chaperone ProQ in the Gram-negative bacterium Pasteurella multocida. (#134)

Emily Gulliver 1 2 , Brandon Sy 3 , Julia Wong 3 , Deanna Deveson Lucas 1 , Ralf Schittenhelm 4 , David Powell 5 , Marina Harper 1 , Jai Tree 3 , John Boyce 1
  1. Infection and Immunity Program, Monash Biomedicine Discovery Institute and Department of Microbiology, Monash University, Clayton, Vic, Australia
  2. Centre for Innate Immunity and Infectious Diseases, Hudson Institute of Medical Research, Clayton, Victoria, Australia
  3. School of Biotechnology and Biomolecular Sciences, University of New South Wales, Sydney, NSW, Australia
  4. Monash Biomedical Proteomics Facility, Monash Biomedicine Discovery Institute and Department of Biochemistry and Molecular Biology, Monash University , Clayton, Vic, Australia
  5. Monash Bioinformatics Platform, Monash University, Clayton, VIC, Australia

Pasteurella multocida is the causative agent of many diseases, including fowl cholera in chickens. Riboregulation is an important mechanism by which bacteria regulate transcript abundance and protein production through small RNA (sRNA) molecules binding to mRNA molecules with a protein chaperone. Recently, a sRNA chaperone called ProQ has been shown to play a critical role in stabilizing some sRNA/mRNA interactions. To assess the role of ProQ in the Gram-negative bacterium P. multocida, we analyzed the transcriptome of a proQ TargeTron® mutant using RNA‑seq and we used UV- crosslinking, ligation and sequencing of hybrids (UV-CLASH) to identify RNA species that directly bind to ProQ. The RNA-seq identified 43 genes with increased production in the proQ mutant and 104 genes with decreased expression, indicating that ProQ mainly acts to stabilize transcripts. Of the total 147 genes that were differentially expressed, 14 were tRNA genes and of these, seven were identified as binding to ProQ. There were also 24 sRNAs identified through these analyses as either being differentially expressed in a proQ mutant or as being bound by ProQ. From the 24 sRNAs, the Prrc13 sRNA was further characterized and it was determined through Northern blotting that ProQ acts to stabilize the Prrc13 transcript. UV-CLASH also identified three putative mRNA targets of Prrc13, the most notable being adk which is an essential gene in P. multocida strain VP161, indicating that Prrc13 may be an ideal candidate for designing novel therapeutics against P. multocida.