title: Import of the transfer RNase colicin D requires site-specific interaction with the energy-transducing protein TonB.
abstract: The transfer RNase colicin D and ionophoric colicin B appropriate the outer membrane iron siderophore receptor FepA and share a common translocation requirement for the TonB pathway to cross the outer membrane. Despite the almost identical sequences of the N-terminal domains required for the translocation of colicins D and B, two spontaneous tonB mutations (Arg158Ser and Pro161Leu) completely abolished colicin D toxicity but did not affect either the sensitivity to other colicins or the FepA-dependent siderophore uptake capacity. The sensitivity to colicin D of both tonB mutants was fully restored by specific suppressor mutations in the TonB box of colicin D, at Ser18(Thr) and Met19(Ile), respectively. This demonstrates that the interaction of colicin D with TonB is critically dependent on certain residues close to position 160 in TonB and on the side chains of certain residues in the TonB box of colicin D. The effect of introducing the TonB boxes from other TonB-dependent receptors and colicins into colicins D and B was studied. The results of these and other changes in the two TonB boxes show that the role of residues at positions 18 and 19 in colicin D is strongly modulated by other nearby and/or distant residues and that the overall function of colicin D is much more dependent on the interaction with TonB involving the TonB box than is the function of colicin B.
authors.affiliation: IBPC, CNRS, UPR 9073, 13 rue Pierre et Marie Curie, 75005 Paris, France.
authors.forename: Liliana
authors.initials: L
authors.lastname: Mora
authors.forename: Nancy
authors.initials: N
authors.lastname: Diaz
authors.forename: Richard H
authors.initials: RH
authors.lastname: Buckingham
authors.forename: Miklos
authors.initials: M
authors.lastname: de Zamaroczy
mesh_terms: D001426:Bacterial Proteins / Q000378:metabolism* / Q000502:physiology; D003087:Colicins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D008565:Membrane Proteins / Q000378:metabolism* / Q000502:physiology*; D017354:Point Mutation
chemical_list: D001426:Bacterial Proteins; D003087:Colicins; D008565:Membrane Proteins; C057616:tonB protein, Bacteria