Host derived inflammatory phospholipids regulate rahU (PA0122) gene, protein, and biofilm formation in Pseudomonas aeruginosa |
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| Authors: | Rao Jayasimha DiGiandomenico Antonio Artamonov Mykhaylo Leitinger Norbert Amin Ashok R Goldberg Joanna B |
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| Affiliation: | aResearch Department, Carilion Clinic and Virginia Tech Carilion School of Medicine, Roanoke, VA 24013, USA;bDepartment of Bio-Medical Engineering, Virginia Tech and Virginia College of Osteopathic Medicne, Blacksburg, VA 24061, USA;cWake Forest University, Winston-Salem, NC 27109, USA;dCardiovascular Research Center, University of Virginia Health Sciences Center, Charlottesville, VA 22908, USA;eDepartment of Microbiology, University of Virginia Health Sciences Center, Charlottesville, VA 22908, USA |
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| Abstract: | This study describes the role of “inflammatory” oxidized (Ox) phospholipids in regulation of rahU (PA0122) expression and biofilm formation in Pseudomonas aeruginosa (383) wild type (rahU+) and rahU mutant (rahU−) strains. Functional analysis of RahU protein from P. aeruginosa in presence of Ox-phospholipids show: (a) LysoPC modulates RahU gene/and protein expression in rahU+ cells; (b) rahU promoter activity is increased by lysoPC and inhibited by PAPC, Ox-PAPC and arachidonic acid; the latter inhibitory effect can be reversed by lysoPC, which was enzymatically derived from PAPC; (c) biofilm formation increased in rahU− cells as compared to rahU+; and (d) inhibition of rahU promoter activity by PAPC and AA (but not lysoPC) showed significantly augmented biofilm formation in rahU+ but not in rahU− cells. This study shows that host derived Ox-phospholipids affect P. aeruginosa-rahU gene and protein expression, which in turn modulates biofilm formation. The accompanying paper describes the role of RahU protein in eukaryotic-host cells. |
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| Keywords: | Abbreviations: Wild type P. aeruginosa strain 383, expressing PA0122 protein (rahU+) Mutant P. aeruginosa strain 383, lacking expression of PA0122 protein (rahU&minus ) LysoPC, lysophosphotidylcholine, 1-hexanoyl-2-hydroxy-sn-glycero- 3-phosphocholine PAPC, 1-palmitoyl-2-arachidonoyl-sn-phosphatidylcholine Ox-PAPC, oxidized-1-palmitoyl-2-arachidonoyl-sn-phosphatidylcholine PLA2, phospholipase A2 AA, arachidonic acid Gm, gentamicin Tc, tetracycline Sp, spectinomycin Cb, Carbenicillin TMB, 3,3&prime ,5,5&prime -tetramethylbenzidine CV, crystal violet TSA, tryptic soya agar plate BA, blood agar plates LB, Luria&ndash Bertani medium RahU, a Sanskrit word for &ldquo the seizer&rdquo Asp-hemolysin, Aspergillus-hemolysinprotein BHT, Butylated hydroxytoluene |
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