title: HipA-mediated antibiotic persistence via phosphorylation of the glutamyl-tRNA-synthetase.
abstract: Bacterial persistence has been shown to be an underlying factor in the failure of antibiotic treatments. Although many pathways, among them the stringent response and toxin-antitoxin modules, have been linked to antibiotic persistence, a clear molecular mechanism for the growth arrest that characterizes persistent bacteria remained elusive. Here, we screened an expression library for putative targets of HipA, the first toxin linked to persistence, and a serine/threonine kinase. We found that the expression of GltX, the glutamyl-tRNA-synthetase, reverses the toxicity of HipA and prevents persister formation. We show that upon HipA expression, GltX undergoes phosphorylation at Ser239, its ATP-binding site. This phosphorylation leads to accumulation of uncharged tRNA(Glu) in the cell, which results in the activation of the stringent response. Our findings demonstrate a mechanism for persister formation by the hipBA toxin-antitoxin module and provide an explanation for the long-observed connection between persistence and the stringent response.
authors.affiliation: Department of Developmental Biology and Cancer Research, Institute for Medical Research Israel-Canada (IMRIC), The Hebrew University of Jerusalem, Jerusalem 91120, Israel.
authors.forename: Ilana
authors.initials: I
authors.lastname: Kaspy
authors.affiliation: Racah Institute of Physics and the Center for Nanoscience and Nanotechnology, Edmond J. Safra Campus, The Hebrew University, Jerusalem 91904, Israel.
authors.forename: Eitan
authors.initials: E
authors.lastname: Rotem
authors.affiliation: Racah Institute of Physics and the Center for Nanoscience and Nanotechnology, Edmond J. Safra Campus, The Hebrew University, Jerusalem 91904, Israel.
authors.forename: Noga
authors.initials: N
authors.lastname: Weiss
authors.affiliation: Racah Institute of Physics and the Center for Nanoscience and Nanotechnology, Edmond J. Safra Campus, The Hebrew University, Jerusalem 91904, Israel.
authors.forename: Irine
authors.initials: I
authors.lastname: Ronin
authors.affiliation: Racah Institute of Physics and the Center for Nanoscience and Nanotechnology, Edmond J. Safra Campus, The Hebrew University, Jerusalem 91904, Israel.
authors.forename: Nathalie Q
authors.initials: NQ
authors.lastname: Balaban
authors.affiliation: Department of Developmental Biology and Cancer Research, Institute for Medical Research Israel-Canada (IMRIC), The Hebrew University of Jerusalem, Jerusalem 91120, Israel.
authors.forename: Gad
authors.initials: G
authors.lastname: Glaser
mesh_terms: D000255:Adenosine Triphosphate / Q000737:chemistry; D000900:Anti-Bacterial Agents / Q000494:pharmacology; D001426:Bacterial Proteins / Q000378:metabolism; D001665:Binding Sites; D003001:Cloning, Molecular; D024881:Drug Resistance, Bacterial / Q000235:genetics*; D004926:Escherichia coli / Q000187:drug effects / Q000378:metabolism*; D029968:Escherichia coli Proteins / Q000378:metabolism*; D015964:Gene Expression Regulation, Bacterial; D015723:Gene Library; D005975:Glutamate-tRNA Ligase / Q000378:metabolism*; D010641:Phenotype; D010766:Phosphorylation; D012694:Serine / Q000378:metabolism; D013997:Time Factors
chemical_list: D000900:Anti-Bacterial Agents; D001426:Bacterial Proteins; D029968:Escherichia coli Proteins; C070741:hipA protein, E coli; D012694:Serine; D000255:Adenosine Triphosphate; D005975:Glutamate-tRNA Ligase