title: A Novel Phosphodiesterase of the GdpP Family Modulates Cyclic di-AMP Levels in Response to Cell Membrane Stress in Daptomycin-Resistant Enterococci.
abstract: Substitutions in the LiaFSR membrane stress pathway are frequently associated with the emergence of antimicrobial peptide resistance in bothEnterococcus faecalisandEnterococcus faeciumCyclic di-AMP (c-di-AMP) is an important signal molecule that affects many aspects of bacterial physiology, including stress responses. We have previously identified a mutation in a gene (designatedyybT) inE. faecalisthat was associated with the development of daptomycin resistance, resulting in a change at position 440 (yybTI440S) in the predicted protein. Here, we show that intracellular c-di-AMP signaling is present in enterococci, and on the basis ofin vitrophysicochemical characterization, we show thatE. faecalisyybTencodes a cyclic dinucleotide phosphodiesterase of the GdpP family that exhibits specific activity toward c-di-AMP by hydrolyzing it to 5'pApA. TheE. faecalisGdpPI440Ssubstitution reduces c-di-AMP phosphodiesterase activity more than 11-fold, leading to further increases in c-di-AMP levels. Additionally, deletions ofliaR(encoding the response regulator of the LiaFSR system) that lead to daptomycin hypersusceptibility in bothE. faecalisandE. faeciumalso resulted in increased c-di-AMP levels, suggesting that changes in the LiaFSR stress response pathway are linked to broader physiological changes. Taken together, our data show that modulation of c-di-AMP pools is strongly associated with antibiotic-induced cell membrane stress responses via changes in GdpP activity or signaling through the LiaFSR system.
authors.affiliation: Department of Biosciences, Rice University, Houston, Texas, USA.
authors.forename: Xu
authors.identifier: 0000-0003-1309-1848
authors.initials: X
authors.lastname: Wang
authors.affiliation: Department of Biosciences, Rice University, Houston, Texas, USA.
authors.forename: Milya
authors.initials: M
authors.lastname: Davlieva
authors.affiliation: Molecular Genetics and Antimicrobial Resistance Unit, International Center for Microbial Genomics, Universidad El Bosque, Bogota, Colombia.
authors.forename: Jinnethe
authors.initials: J
authors.lastname: Reyes
authors.affiliation: Center for Antimicrobial Resistance and Microbial Genomics, Division of Infectious Diseases, University of Texas McGovern School of Medicine at Houston, Houston, Texas, USA.|Molecular Genetics and Antimicrobial Resistance Unit, International Center for Microbial Genomics, Universidad El Bosque, Bogota, Colombia.
authors.forename: Diana
authors.initials: D
authors.lastname: Panesso
authors.affiliation: Center for Antimicrobial Resistance and Microbial Genomics, Division of Infectious Diseases, University of Texas McGovern School of Medicine at Houston, Houston, Texas, USA.|Department of Microbiology and Molecular Genetics, University of Texas McGovern School of Medicine at Houston, Houston, Texas, USA.|Molecular Genetics and Antimicrobial Resistance Unit, International Center for Microbial Genomics, Universidad El Bosque, Bogota, Colombia.
authors.forename: Cesar A
authors.initials: CA
authors.lastname: Arias
authors.affiliation: Department of Biosciences, Rice University, Houston, Texas, USA Shamoo@rice.edu.
authors.forename: Yousif
authors.initials: Y
authors.lastname: Shamoo
mesh_terms: D020816:Amino Acid Motifs; D000900:Anti-Bacterial Agents / Q000494:pharmacology; D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D001665:Binding Sites; D002462:Cell Membrane / Q000187:drug effects* / Q000378:metabolism; D003001:Cloning, Molecular; D017576:Daptomycin / Q000494:pharmacology; D015226:Dinucleoside Phosphates / Q000378:metabolism*; D024881:Drug Resistance, Bacterial / Q000235:genetics; D013293:Enterococcus faecalis / Q000187:drug effects* / Q000235:genetics / Q000378:metabolism; D016984:Enterococcus faecium / Q000187:drug effects* / Q000235:genetics / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D015964:Gene Expression Regulation, Bacterial; D007700:Kinetics; D008826:Microbial Sensitivity Tests; D008958:Models, Molecular; D009154:Mutation; D010727:Phosphoric Diester Hydrolases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D011485:Protein Binding; D000072756:Protein Conformation, alpha-Helical; D000072757:Protein Conformation, beta-Strand; D054730:Protein Interaction Domains and Motifs; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D015398:Signal Transduction; D013312:Stress, Physiological; D013379:Substrate Specificity
chemical_list: D000900:Anti-Bacterial Agents; D001426:Bacterial Proteins; D015226:Dinucleoside Phosphates; D011994:Recombinant Proteins; C528998:cyclic diadenosine phosphate; D010727:Phosphoric Diester Hydrolases; D017576:Daptomycin
keywords: Enterococcus faecalis; Enterococcus faecium; GdpP; LiaFSR; c-di-AMP; cyclic dinucleotide; daptomycin; enterococci; membrane stress