Transposon mutagenesis screen in Klebsiella pneumoniae identifies genetic determinants required for growth in human urine and serum

• Published in: eLife Vol. 12
• Main Authors: Gray, Jessica, Torres, Von Vergel L, Goodall, Emily, McKeand, Samantha A, Scales, Danielle, Collins, Christy, Wetherall, Laura, Lian, Zheng Jie, Bryant, Jack A, Milner, Matthew T, Dunne, Karl A, Icke, Christopher, Rooke, Jessica L, Schneiders, Thamarai, Lund, Peter A, Cunningham, Adam F, Cole, Jeff A, Henderson, Ian R
• Format: Journal Article
• Language: English
• Published: England eLife Science Publications, Ltd 27.08.2024; eLife Sciences Publications Ltd; eLife Sciences Publications, Ltd
• Subjects: Antimicrobial agents; Bacterial pneumonia; Canada; Chromosomes; DNA Transposable Elements - genetics; Drug resistance in microorganisms; Genes; Genetics and Genomics; genome; Genomes; Genomics; Gram-negative bacteria; Growth; Health aspects; Humans; Infections; Klebsiella; Klebsiella Infections - microbiology; Klebsiella Infections - urine; Klebsiella pneumoniae; Klebsiella pneumoniae - genetics; Klebsiella pneumoniae - growth & development; Microbiology and Infectious Disease; Molecular modelling; Mortality; Multidrug resistance; Mutagenesis; Mutagenesis, Insertional; Phylogenetics; Pneumonia; Public health; serum; Serum - microbiology; TnSeq; TraDIS; Transposon mutagenesis; Transposons; Urine; Urine - microbiology; Urogenital system; Virulence; Virulence (Microbiology); Canada; Klebsiella; TraDIS; genome; genetics; infectious disease; urine; TnSeq; microbiology; genomics; serum
• ISSN: 2050-084X; 2050-084X
• DOI: 10.7554/eLife.88971

is a global public health concern due to the rising myriad of hypervirulent and multidrug-resistant clones both alarmingly associated with high mortality. The molecular mechanisms underpinning these recalcitrant infection, and how virulence is coupled with the emergence of lineages resistant to nearly all present-day clinically important antimicrobials, are unclear. In this study, we performed a genome-wide screen in ECL8, a member of the endemic K2-ST375 pathotype most often reported in Asia, to define genes essential for growth in a nutrient-rich laboratory medium (Luria-Bertani [LB] medium), human urine, and serum. Through transposon directed insertion-site sequencing (TraDIS), a total of 427 genes were identified as essential for growth on LB agar, whereas transposon insertions in 11 and 144 genes decreased fitness for growth in either urine or serum, respectively. These studies not only provide further knowledge on the genetics of this pathogen but also provide a strong impetus for discovering new antimicrobial targets to improve current therapeutic options for infections.