Gene expression of Pseudomonas aeruginosa in a mucin-containing synthetic growth medium mimicking cystic fibrosis lung sputum.
Pseudomonas aeruginosa airway infection is the leading cause of morbidity and mortality in cystic fibrosis (CF) patients. Various in vitro models have been developed to study P. aeruginosa pathobiology in the CF lung. In this study we produced a modified artificial-sputum medium (ASMDM) more closely resembling CF sputum than previous models, and extended previous work by using strain PAO1 arrays to examine the global transcription profiles of P. aeruginosa strain UCBPP-PA14 under early exponential-phase and stationary-phase growth. In early exponential phase, 38/39 nutrition-related genes were upregulated in line with data from previous in vitro models using UCBPP-PA14. Additionally, 23 type III secretion system (T3SS) genes, several anaerobic respiration genes and 24 quorum-sensing (QS)-related genes were upregulated in ASMDM, suggesting enhanced virulence factor expression and priming for anaerobic growth and biofilm formation. Under stationary phase growth in ASMDM, macroscopic clumps resembling microcolonies were evident in UCBPP-PA14 and CF strains, and over 40 potentially important genes were differentially expressed relative to stationary-phase growth in Luria broth. Most notably, QS-related and T3SS genes were downregulated in ASMDM, and iron-acquisition and assimilatory nitrate reductase genes were upregulated, simulating the iron-depleted, microaerophilic/anaerobic environment of CF sputum. ASMDM thus appears to be highly suitable for gene expression studies of P. aeruginosa in CF.
Fung
Carina
C
Department of Infectious Diseases and Immunology, University of Sydney, Sydney, Australia.
Naughton
Sharna
S
Department of Infectious Diseases and Immunology, University of Sydney, Sydney, Australia.
Turnbull
Lynne
L
Institute for the Biotechnology of Infectious Diseases, University of Technology, Sydney, Australia.
Tingpej
Pholawat
P
Department of Infectious Diseases and Immunology, University of Sydney, Sydney, Australia.
Rose
Barbara
B
Department of Infectious Diseases and Immunology, University of Sydney, Sydney, Australia.
Arthur
Jonathan
J
Sydney Bioinformatics, University of Sydney, Sydney, Australia.|Discipline of Medicine, Sydney Medical School, University of Sydney, Sydney, Australia.
Hu
Honghua
H
Department of Infectious Diseases and Immunology, University of Sydney, Sydney, Australia.
Harmer
Christopher
C
Department of Infectious Diseases and Immunology, University of Sydney, Sydney, Australia.
Harbour
Colin
C
Department of Infectious Diseases and Immunology, University of Sydney, Sydney, Australia.
Hassett
Daniel J
DJ
Department of Molecular Genetics, Biochemistry and Microbiology, University of Cincinnati College of Medicine, Cincinnati, OH, USA.
Whitchurch
Cynthia B
CB
Institute for the Biotechnology of Infectious Diseases, University of Technology, Sydney, Australia.
Manos
Jim
J
Department of Infectious Diseases and Immunology, University of Sydney, Sydney, Australia.
D003470:Culture Media / Q000737:chemistry* / Q000494:pharmacology*; D003550:Cystic Fibrosis / Q000503:physiopathology; D020869:Gene Expression Profiling; D015964:Gene Expression Regulation, Bacterial / Q000187:drug effects*; D006801:Humans; D009077:Mucins / Q000737:chemistry* / Q000494:pharmacology*; D011550:Pseudomonas aeruginosa / Q000502:physiology*; D013183:Sputum
D003470:Culture Media; D009077:Mucins