title: Molecular basis for erythromycin-dependent ribosome stalling during translation of the ErmBL leader peptide.
abstract: In bacteria, ribosome stalling during translation of ErmBL leader peptide occurs in the presence of the antibiotic erythromycin and leads to induction of expression of the downstream macrolide resistance methyltransferase ErmB. The lack of structures of drug-dependent stalled ribosome complexes (SRCs) has limited our mechanistic understanding of this regulatory process. Here we present a cryo-electron microscopy structure of the erythromycin-dependent ErmBL-SRC. The structure reveals that the antibiotic does not interact directly with ErmBL, but rather redirects the path of the peptide within the tunnel. Furthermore, we identify a key peptide-ribosome interaction that defines an important relay pathway from the ribosomal tunnel to the peptidyltransferase centre (PTC). The PTC of the ErmBL-SRC appears to adopt an uninduced state that prevents accommodation of Lys-tRNA at the A-site, thus providing structural basis for understanding how the drug and the nascent peptide cooperate to inhibit peptide bond formation and induce translation arrest.
authors.affiliation: Gene Center, Department for Biochemistry, University of Munich, Feodor-Lynen Strasse 25, 81377 Munich, Germany.
authors.forename: Stefan
authors.initials: S
authors.lastname: Arenz
authors.affiliation: Center for Pharmaceutical Biotechnology, University of Illinois, Chicago, Illinois 60607, USA.
authors.forename: Haripriya
authors.initials: H
authors.lastname: Ramu
authors.affiliation: Center for Pharmaceutical Biotechnology, University of Illinois, Chicago, Illinois 60607, USA.
authors.forename: Pulkit
authors.initials: P
authors.lastname: Gupta
authors.affiliation: Gene Center, Department for Biochemistry, University of Munich, Feodor-Lynen Strasse 25, 81377 Munich, Germany.
authors.forename: Otto
authors.initials: O
authors.lastname: Berninghausen
authors.affiliation: 1] Gene Center, Department for Biochemistry, University of Munich, Feodor-Lynen Strasse 25, 81377 Munich, Germany [2] Center for integrated Protein Science Munich (CiPSM), University of Munich, Feodor-Lynen Strasse 25, 81377 Munich, Germany.
authors.forename: Roland
authors.initials: R
authors.lastname: Beckmann
authors.affiliation: Center for Pharmaceutical Biotechnology, University of Illinois, Chicago, Illinois 60607, USA.
authors.forename: Nora
authors.initials: N
authors.lastname: Vázquez-Laslop
authors.affiliation: Center for Pharmaceutical Biotechnology, University of Illinois, Chicago, Illinois 60607, USA.
authors.forename: Alexander S
authors.initials: AS
authors.lastname: Mankin
authors.affiliation: 1] Gene Center, Department for Biochemistry, University of Munich, Feodor-Lynen Strasse 25, 81377 Munich, Germany [2] Center for integrated Protein Science Munich (CiPSM), University of Munich, Feodor-Lynen Strasse 25, 81377 Munich, Germany.
authors.forename: Daniel N
authors.initials: DN
authors.lastname: Wilson
mesh_terms: D000595:Amino Acid Sequence; D001483:Base Sequence; D020285:Cryoelectron Microscopy; D004917:Erythromycin / Q000494:pharmacology*; D015964:Gene Expression Regulation, Bacterial / Q000187:drug effects*; D008780:Methyltransferases / Q000235:genetics*; D008958:Models, Molecular; D008969:Molecular Sequence Data; D009841:Oligonucleotides / Q000235:genetics; D014176:Protein Biosynthesis / Q000187:drug effects*; D021382:Protein Sorting Signals / Q000235:genetics*; D012270:Ribosomes / Q000187:drug effects / Q000502:physiology*
chemical_list: D009841:Oligonucleotides; D021382:Protein Sorting Signals; D004917:Erythromycin; D008780:Methyltransferases; C035181:rRNA (adenosine-O-2'-)methyltransferase