Genome-Wide High-Throughput Screening to Investigate Essential Genes Involved in Methicillin-Resistant Staphylococcus aureus Sequence Type 398 Survival

Mette Theilgaard Christiansen, Rolf Sommer Kaas, Roy R. Chaudhuri, Mark A. Holmes, Henrik Hasman, Frank Møller Aarestrup

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Abstract

Livestock-associated methicillin-resistant Staphylococcus aureus (LA-MRSA) Sequence Type 398 (ST398) is an opportunistic pathogen that is able to colonize and cause disease in several animal species including humans. To better understand the adaptation, evolution, transmission and pathogenic capacity, further investigations into the importance of the different genes harboured by LA-MRSA ST398 are required. In this study we generated a genome-wide transposon mutant library in an LA-MRSA ST398 isolate to evaluate genes important for bacterial survival in laboratory and host-specific environments. The transposon mutant library consisted of approximately 1 million mutants with around 140,000 unique insertion sites and an average number of unique inserts per gene of 44.8. We identified LA-MRSA ST398 essential genes comparable to other high-throughput S. aureus essential gene studies. As ST398 is the most common MRSA isolated from pigs, the transposon mutant library was screened in whole porcine blood. Twenty-four genes were specifically identified as important for bacterial survival in porcine blood. Mutations in 23 of these genes resulted in attenuated bacterial fitness. Seven of the 23 genes were of unknown function, whereas 16 genes were annotated with functions predominantly related to carbon metabolism, pH shock and a variety of regulations and only indirectly to virulence factors. Mutations in one gene of unknown function resulted in a hypercompetitive mutant. Further evaluation of these genes is required to determine their specific relevance in blood survival.
Original languageEnglish
Article numbere89018
JournalPLOS ONE
Volume9
Issue number2
Number of pages13
ISSN1932-6203
DOIs
Publication statusPublished - 2014

Keywords

  • Artiodactyla Mammalia Vertebrata Chordata Animalia (Animals, Artiodactyls, Chordates, Mammals, Nonhuman Vertebrates, Nonhuman Mammals, Vertebrates) - Suidae [85740] pig common
  • Gram-Positive Cocci Eubacteria Bacteria Microorganisms (Bacteria, Eubacteria, Microorganisms) - Micrococcaceae [07702] Staphylococcus aureus species pathogen
  • carbon 7440-44-0 metabolism
  • gene mutation
  • methicillin 61-32-5 antibacterial-drug, antiinfective-drug
  • 03502, Genetics - General
  • 03506, Genetics - Animal
  • 10060, Biochemistry studies - General
  • 10062, Biochemistry studies - Nucleic acids, purines and pyrimidines
  • 12502, Pathology - General
  • 12512, Pathology - Therapy
  • 15002, Blood - Blood and lymph studies
  • 15004, Blood - Blood cell studies
  • 22002, Pharmacology - General
  • 31000, Physiology and biochemistry of bacteria
  • 31500, Genetics of bacteria and viruses
  • 36002, Medical and clinical microbiology - Bacteriology
  • 38502, Chemotherapy - General, methods and metabolism
  • 38504, Chemotherapy - Antibacterial agents
  • Infection
  • Methods and Techniques
  • Molecular Genetics
  • Pharmacology
  • drug resistance
  • Staphylococcus aureus infection bacterial disease pathology, mortality, genetics, transmission
  • Biochemistry and Molecular Biophysics
  • blood blood and lymphatics
  • AM990992 GenBank, EMBL, DDJB nucleotide sequence
  • genome-wide high-throuhput screening laboratory techniques, genetic techniques
  • MULTIDISCIPLINARY
  • STATIONARY-PHASE
  • VIRULENCE GENES
  • MUTAGENESIS
  • EXPRESSION
  • PEPTIDOGLYCAN
  • STRAINS
  • PROTEIN
  • IDENTIFICATION
  • GROWTH
  • ACID

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