title: Identification of a target cell permissive factor required for contact-dependent growth inhibition (CDI).
abstract: Bacterial contact-dependent growth inhibition (CDI) is mediated by the CdiB/CdiA family of two-partner secretion proteins. CdiA effector proteins are exported onto the surface of CDI(+) inhibitor cells, where they interact with susceptible bacteria and deliver effectors/toxins derived from their C-terminal regions (CdiA-CT). CDI(+) cells also produce an immunity protein that binds the CdiA-CT and blocks its activity to prevent autoinhibition. Here, we show that the CdiA-CT from uropathogenic Escherichia coli strain 536 (UPEC536) is a latent tRNase that requires activation by the biosynthetic enzyme CysK (O-acetylserine sulfhydrylase A). UPEC536 CdiA-CT exhibits no nuclease activity in vitro, but cleaves within transfer RNA (tRNA) anti-codon loops when purified CysK is added. CysK and CdiA-CT form a stable complex, and their binding interaction appears to mimic that of the CysK/CysE cysteine synthase complex. CdiA-CT activation is also required for growth inhibition. Synthesis of CdiA-CT in E. coli cysK(+) cells arrests cell growth, whereas the growth of ΔcysK mutants is unaffected by the toxin. Moreover, E. coli ΔcysK cells are completely resistant to inhibitor cells expressing UPEC536 CdiA, indicating that CysK is required to activate the tRNase during CDI. Thus, CysK acts as a permissive factor for CDI, providing a potential mechanism to modulate growth inhibition in target cells.
authors.affiliation: Department of Molecular, Cellular, and Developmental Biology, University of California at Santa Barbara, CA 93106, USA.
authors.forename: Elie J
authors.initials: EJ
authors.lastname: Diner
authors.forename: Christina M
authors.initials: CM
authors.lastname: Beck
authors.forename: Julia S
authors.initials: JS
authors.lastname: Webb
authors.forename: David A
authors.initials: DA
authors.lastname: Low
authors.forename: Christopher S
authors.initials: CS
authors.lastname: Hayes
mesh_terms: D000595:Amino Acid Sequence; D003067:Coenzymes / Q000378:metabolism; D003260:Contact Inhibition / Q000235:genetics; D003547:Cysteine Synthase / Q000235:genetics / Q000378:metabolism; D004789:Enzyme Activation; D004926:Escherichia coli / Q000201:enzymology* / Q000254:growth & development*; D029968:Escherichia coli Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D008565:Membrane Proteins / Q000737:chemistry / Q000378:metabolism*; D008969:Molecular Sequence Data; D011485:Protein Binding; D016415:Sequence Alignment
chemical_list: C503242:CdiA protein, E coli; D003067:Coenzymes; D029968:Escherichia coli Proteins; D008565:Membrane Proteins; D003547:Cysteine Synthase