Chemical Communication of Antibiotic Resistance by a Highly Resistant Subpopulation of Bacterial Cells.docVIP

ChemicalCommunicationofAntibioticResistancebya HighlyResistantSubpopulationofBacterialCells OmarM.El-Halfawy1,2,MiguelA.Valvano1,3 * 1Centre for Human Immunology and Department of Microbiology and Immunology, University of Western Ontario, London, Ontario, Canada, 2Department of Pharmaceutical Microbiology, Faculty of Pharmacy, Alexandria University, Alexandria, Egypt, 3Centre for Infection and Immunity, Queen’s University Belfast, Belfast, UnitedKingdom Abstract Theoverallantibioticresistanceofabacterialpopulationresultsfromthecombinationofawiderangeofsusceptibilities displayed by subsets of bacterial cells. Bacterial heteroresistance to antibiotics has been documented for several opportunisticGram-negativebacteria,butthemechanismofheteroresistanceisunclear.WeuseBurkholderiacenocepacia asamodelopportunisticbacteriumtoinvestigatetheimplicationsofheterogeneityintheresponsetotheantimicrobial peptidepolymyxinB(PmB)andalsootherbactericidalantibiotics.Here,wereportthatB.cenocepaciaisheteroresistantto PmB. Population analysis profiling also identified B. cenocepacia subpopulations arising from a seemingly homogenous culturethatareresistanttohigherlevelsofpolymyxinBthantherestofthecellsintheculture,andcanprotectthemore sensitivecellsfromkilling,aswellassensitivebacteriafromotherspecies,suchasPseudomonasaeruginosaandEscherichia coli. Communication of resistance depended on upregulation of putrescine synthesis and YceI, a widely conserved low- molecularweightsecretedprotein.DeletionofgenesforthesynthesisofputrescineandYceIabrogateprotection,while pharmacologic inhibition of putrescine synthesis reduced resistance to polymyxin B. Polyamines and YceI were also required for heteroresistance of B. cenocepacia to various bactericidal antibiotics. We propose that putrescine and YceI resemble danger infochemicals whose increased production by a bacterial subpopulation, becoming more resistant to bactericidalantibiotics,communicateshigherlevelofresistancetomoresensitivemembersofth