The evolution of antimicrobial peptide resistance in Pseudomonas aeruginosa is shaped by strong epistatic interactions.
Colistin is an antimicrobial peptide that has become the only remaining alternative for the treatment of multidrug-resistant Gram-negative bacterial infections, but little is known of how clinical levels of colistin resistance evolve. We use in vitro experimental evolution and whole-genome sequencing of colistin-resistant Pseudomonas aeruginosa isolates from cystic fibrosis patients to reconstruct the molecular evolutionary pathways open for high-level colistin resistance. We show that the evolution of resistance is a complex, multistep process that requires mutation in at least five independent loci that synergistically create the phenotype. Strong intergenic epistasis limits the number of possible evolutionary pathways to resistance. Mutations in transcriptional regulators are essential for resistance evolution and function as nodes that potentiate further evolution towards higher resistance by functionalizing and increasing the effect of the other mutations. These results add to our understanding of clinical antimicrobial peptide resistance and the prediction of resistance evolution.
Department of Systems Biology, Technical University of Denmark, Kongens Lyngby, Denmark.
Nicholas
N
Jochumsen
Department of Systems Biology, Technical University of Denmark, Kongens Lyngby, Denmark.|Center for Genomic Medicine, Rigshospitalet, 2100 Copenhagen, Denmark.
Rasmus L
http://orcid.org/0000-0002-5267-3173
RL
Marvig
Department of Systems Biology, Technical University of Denmark, Kongens Lyngby, Denmark.
Søren
S
Damkiær
Department of Systems Biology, Technical University of Denmark, Kongens Lyngby, Denmark.
Rune Lyngklip
RL
Jensen
Department of Veterinary Disease Biology, University of Copenhagen, 1870 Frederiksberg C, Denmark.
Wilhelm
W
Paulander
Department of Systems Biology, Technical University of Denmark, Kongens Lyngby, Denmark.
Søren
S
Molin
Department of Systems Biology, Technical University of Denmark, Kongens Lyngby, Denmark.
Lars
L
Jelsbak
National Veterinary Institute, Technical University of Denmark, Frederiksberg, Denmark.
Anders
http://orcid.org/0000-0002-2175-830X
A
Folkesson
D000483:Alleles; D023181:Antimicrobial Cationic Peptides / Q000737:chemistry*; D003091:Colistin / Q000737:chemistry*; D003550:Cystic Fibrosis / Q000382:microbiology; D004252:DNA Mutational Analysis; D024881:Drug Resistance, Bacterial; D004843:Epistasis, Genetic; D004926:Escherichia coli / Q000378:metabolism; D019143:Evolution, Molecular; D016681:Genome, Fungal; D009154:Mutation; D010455:Peptides / Q000737:chemistry; D010641:Phenotype; D020641:Polymorphism, Single Nucleotide; D011550:Pseudomonas aeruginosa / Q000235:genetics* / Q000378:metabolism*
D023181:Antimicrobial Cationic Peptides; D010455:Peptides; D003091:Colistin
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