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Charlotte Rohde Knudsen

Aminoacyl-tRNA-charged eukaryotic elongation factor 1A is a bona fide substrate for Legionelle pneumophila effector glucosyltransferases

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DOI

  • Tina Tzivelekidis, Albert-Ludwigs-Universität, Freiburg, Tyskland
  • Thomas Jank, Albert-Ludwigs-Universität, Freiburg, Tyskland
  • Corinna Pohl, Max-Planck-Institut für biophysikalische Chemie, Göttingen, Tyskland
  • Andreas Schlosser, Albert-Ludwigs-Universität, Freiburg, Tyskland
  • Sabine Rospert, Albert-Ludwigs-Universität, Freiburg, Tyskland
  • Charlotte Rohde Knudsen
  • Marina Rodnina, Max-Planck-Institut für biophysikalische Chemie, Göttingen, Tyskland
  • Yuri Belyi, Gamaleya Research Institute, Moscow, Rusland
  • Klaus Aktories, Albert-Ludwigs-Universität, Freiburg, Tyskland
Legionella pneumophila, which is the causative organism of Legionnaires disease, translocates numerous effector proteins into the host cell cytosol by a type IV secretion system during infection. Among the most potent effector proteins of Legionella are glucosyltransferases (Lgt’s), which selectively modify eukaryotic elongation factor (eEF) 1A at Ser-53 in the GTP binding domain. Glucosylation results in inhibition of protein synthesis. Here we show that in vitro glucosylation of yeast and mouse eEF1A by Lgt3 in the presence of the factors Phe-tRNAPhe and GTP was enhanced 150 and 590-fold, respectively. The glucosylation of eEF1A catalyzed by Lgt1 and 2 was increased about 70-fold. By comparison of uncharged tRNA with two distinct aminoacyl-tRNAs (His-tRNAHis and Phe-tRNAPhe) we could show that aminoacylation is crucial for Lgtcatalyzed glucosylation. Aminoacyl-tRNA had no effect on the enzymatic properties of Lgt’s and did not enhance the glucosylation rate of eEF1A truncation mutants, consisting of the GTPase domain only or of a 5 kDa peptide covering Ser-53 of eEF1A. Furthermore, binding of aminoacyl-tRNA to eEF1A was not altered by glucosylation. Taken together, our data suggest that the ternary complex, consisting of eEF1A, aminoacyl-tRNA and GTP, is the bona fide substrate for Lgt’s.
OriginalsprogEngelsk
TidsskriftP L o S One
Vol/bind6
Nummer12
Sider (fra-til)e29525
Antal sider11
ISSN1932-6203
DOI
StatusUdgivet - 22 dec. 2011

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