Aminoacyl-tRNA-charged eukaryotic elongation factor 1A is the bona fide substrate for Legionella pneumophila effector glucosyltransferases
| Autor: | Tzivelekidis, T., Jank, T., Pohl, C., Schlosser, A., Rospert, S., Knudsen, C., Rodnina, M., Belyi, Y., Aktories, K. |
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| Jazyk: | angličtina |
| Rok vydání: | 2011 |
| Předmět: |
Models
Molecular Bacterial Diseases Glycosylation Gene Expression Legionella lcsh:Medicine Protein Synthesis RNA Transfer Amino Acyl Biochemistry Microbiology Cell Line Legionella pneumophila Substrate Specificity Mice Adenosine Triphosphate Peptide Elongation Factor 1 Tandem Mass Spectrometry Molecular Cell Biology Animals Amino Acid Sequence Cloning Molecular lcsh:Science Biology Microbial Pathogens Enzyme Classes Macrophages lcsh:R Proteins Enzymes Bacterial Pathogens carbohydrates (lipids) Host-Pathogen Interaction Infectious Diseases Glucosyltransferases Biocatalysis Medicine Protein Translation lcsh:Q Chromatography Liquid Research Article |
| Zdroj: | PLoS ONE, Vol 6, Iss 12, p e29525 (2011) PLoS ONE PLoS One |
| ISSN: | 1932-6203 |
| Popis: | Legionella pneumophila, which is the causative organism of Legionnaireś 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-tRNA(Phe) 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-tRNA(His) and Phe-tRNA(Phe)) we could show that aminoacylation is crucial for Lgt-catalyzed 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. |
| Databáze: | OpenAIRE |
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