title: Arginine-rhamnosylation as new strategy to activate translation elongation factor P.
abstract: Ribosome stalling at polyproline stretches is common and fundamental. In bacteria, translation elongation factor P (EF-P) rescues such stalled ribosomes, but only when it is post-translationally activated. In Escherichia coli, activation of EF-P is achieved by (R)-β-lysinylation and hydroxylation of a conserved lysine. Here we have unveiled a markedly different modification strategy in which a conserved arginine of EF-P is rhamnosylated by a glycosyltransferase (EarP) using dTDP-L-rhamnose as a substrate. This is to our knowledge the first report of N-linked protein glycosylation on arginine in bacteria and the first example in which a glycosylated side chain of a translation elongation factor is essential for function. Arginine-rhamnosylation of EF-P also occurs in clinically relevant bacteria such as Pseudomonas aeruginosa. We demonstrate that the modification is needed to develop pathogenicity, making EarP and dTDP-L-rhamnose-biosynthesizing enzymes ideal targets for antibiotic development.
authors: 1] Center for Integrated Protein Science Munich, Ludwig-Maximilians-Universität München, Munich, Germany. [2] Department of Biology I, Microbiology, Ludwig-Maximilians-Universität München, Martinsried, Germany.
authors: Jürgen
authors: J
authors: Lassak
authors: Proteomics and Signal Transduction, Max-Planck Institute of Biochemistry, Martinsried, Germany.
authors: Eva C
authors: EC
authors: Keilhauer
authors: 1] Center for Integrated Protein Science Munich, Ludwig-Maximilians-Universität München, Munich, Germany. [2] Department of Biology I, Microbiology, Ludwig-Maximilians-Universität München, Martinsried, Germany.
authors: Maximilian
authors: M
authors: Fürst
authors: Max Planck Institute for Terrestrial Microbiology, Marburg, Germany.
authors: Kristin
authors: K
authors: Wuichet
authors: Institute for Molecular Bacteriology, Twincore, Centre for Clinical and Experimental Infection Research, a joint venture of the Helmholtz Centre of Infection Research and the Hannover Medical School, Hannover, Germany.
authors: Julia
authors: J
authors: Gödeke
authors: 1] Center for Integrated Protein Science Munich, Ludwig-Maximilians-Universität München, Munich, Germany. [2] Gene Center, Department for Biochemistry, Ludwig-Maximilians-Universität München, Munich, Germany.
authors: Agata L
authors: AL
authors: Starosta
authors: Department of Pharmaceutical Sciences, College of Pharmacy, University of Kentucky, Lexington, Kentucky, USA.
authors: Jhong-Min
authors: JM
authors: Chen
authors: Max Planck Institute for Terrestrial Microbiology, Marburg, Germany.
authors: Lotte
authors: L
authors: Søgaard-Andersen
authors: Department of Pharmaceutical Sciences, College of Pharmacy, University of Kentucky, Lexington, Kentucky, USA.
authors: Jürgen
authors: J
authors: Rohr
authors: 1] Center for Integrated Protein Science Munich, Ludwig-Maximilians-Universität München, Munich, Germany. [2] Gene Center, Department for Biochemistry, Ludwig-Maximilians-Universität München, Munich, Germany.
authors: Daniel N
authors: DN
authors: Wilson
authors: 1] Institute for Molecular Bacteriology, Twincore, Centre for Clinical and Experimental Infection Research, a joint venture of the Helmholtz Centre of Infection Research and the Hannover Medical School, Hannover, Germany. [2] Department of Molecular Bacteriology, Helmholtz Centre for Infection Research, Braunschweig, Germany.
authors: Susanne
authors: S
authors: Häussler
authors: Proteomics and Signal Transduction, Max-Planck Institute of Biochemistry, Martinsried, Germany.
authors: Matthias
authors: M
authors: Mann
authors: 1] Center for Integrated Protein Science Munich, Ludwig-Maximilians-Universität München, Munich, Germany. [2] Department of Biology I, Microbiology, Ludwig-Maximilians-Universität München, Martinsried, Germany.
authors: Kirsten
authors: K
authors: Jung
mesh_terms: D000595:Amino Acid Sequence; D001120:Arginine / Q000737:chemistry*; D001665:Binding Sites; D045744:Cell Line, Tumor; D002853:Chromatography, Liquid; D018360:Crystallography, X-Ray; D004926:Escherichia coli / Q000378:metabolism; D006031:Glycosylation; D016695:Glycosyltransferases / Q000378:metabolism; D006801:Humans; D006900:Hydroxylation; D008239:Lysine / Q000737:chemistry*; D008390:Markov Chains; D013058:Mass Spectrometry; D008958:Models, Molecular; D008969:Molecular Sequence Data; D010445:Peptide Elongation Factors / Q000737:chemistry*; D010455:Peptides / Q000737:chemistry; D010802:Phylogeny; D014176:Protein Biosynthesis; D011550:Pseudomonas aeruginosa / Q000201:enzymology; D012333:RNA, Messenger / Q000378:metabolism; D011994:Recombinant Proteins / Q000737:chemistry; D012210:Rhamnose / Q000737:chemistry*; D012270:Ribosomes / Q000737:chemistry*; D020592:Shewanella / Q000201:enzymology*; D053719:Tandem Mass Spectrometry
chemical_list: D010445:Peptide Elongation Factors; D010455:Peptides; D012333:RNA, Messenger; D011994:Recombinant Proteins; C020746:factor EF-P; D001120:Arginine; D016695:Glycosyltransferases; D008239:Lysine; D012210:Rhamnose