title: Resuscitation ofPseudomonas aeruginosafrom dormancy requires hibernation promoting factor (PA4463) for ribosome preservation.
abstract: Pseudomonas aeruginosabiofilm infections are difficult to treat with antibiotic therapy in part because the biofilms contain subpopulations of dormant antibiotic-tolerant cells. The dormant cells can repopulate the biofilms following alleviation of antibiotic treatments. While dormant, the bacteria must maintain cellular integrity, including ribosome abundance, to reinitiate the de novo protein synthesis required for resuscitation. Here, we demonstrate that theP. aeruginosagene PA4463 [hibernation promoting factor (HPF)], but not the ribosome modulation factor (PA3049), is required for ribosomal RNA preservation during prolonged nutrient starvation conditions. Single-cell-level studies using fluorescence in situ hybridization (FISH) and growth in microfluidic drops demonstrate that, in the absence ofhpf, the rRNA abundances of starved cells decrease to levels that cause them to lose their ability to resuscitate from starvation, leaving intact nondividing cells.P. aeruginosadefective in the stringent response also had reduced ability to resuscitate from dormancy. However, FISH analysis of the starved stringent response mutant showed a bimodal response where the individual cells contained either abundant or low ribosome content, compared with the wild-type strain. The results indicate that ribosome maintenance is key for maintaining the ability ofP. aeruginosato resuscitate from starvation-induced dormancy and that HPF is the major factor associated withP. aeruginosaribosome preservation.
authors.affiliation: Department of Microbiology and Immunology, Montana State University, Bozeman, MT 59717.|Center for Biofilm Engineering, Montana State University, Bozeman, MT 59717.
authors.forename: Tatsuya
authors.initials: T
authors.lastname: Akiyama
authors.affiliation: Department of Microbiology and Immunology, Montana State University, Bozeman, MT 59717.|Center for Biofilm Engineering, Montana State University, Bozeman, MT 59717.
authors.forename: Kerry S
authors.initials: KS
authors.lastname: Williamson
authors.affiliation: Center for Biofilm Engineering, Montana State University, Bozeman, MT 59717.|Department of Chemical and Biological Engineering, Montana State University, Bozeman, MT 59717.
authors.forename: Robert
authors.initials: R
authors.lastname: Schaefer
authors.affiliation: Center for Biofilm Engineering, Montana State University, Bozeman, MT 59717.|Department of Chemical and Biological Engineering, Montana State University, Bozeman, MT 59717.
authors.forename: Shawna
authors.initials: S
authors.lastname: Pratt
authors.affiliation: Center for Biofilm Engineering, Montana State University, Bozeman, MT 59717.|Department of Chemical and Biological Engineering, Montana State University, Bozeman, MT 59717.
authors.forename: Connie B
authors.initials: CB
authors.lastname: Chang
authors.affiliation: Department of Microbiology and Immunology, Montana State University, Bozeman, MT 59717; franklin@montana.edu.|Center for Biofilm Engineering, Montana State University, Bozeman, MT 59717.
authors.forename: Michael J
authors.initials: MJ
authors.lastname: Franklin
mesh_terms: D004734:Energy Metabolism; D015964:Gene Expression Regulation, Bacterial; D006605:Hibernation; D011550:Pseudomonas aeruginosa / Q000502:physiology*; D012335:RNA, Ribosomal / Q000235:genetics / Q000378:metabolism; D012269:Ribosomal Proteins / Q000378:metabolism*; D012270:Ribosomes / Q000378:metabolism*
chemical_list: D012335:RNA, Ribosomal; D012269:Ribosomal Proteins
keywords: dormancy; hibernation promoting factor; rRNA; resuscitation; ribosome