Back to Search
Start Over
Elongation factor 1A is the target of growth inhibition in yeast caused by Legionella pneumophila glucosyltransferase Lgt1
- Source :
- The Journal of biological chemistry. 287(31)
- Publication Year :
- 2012
-
Abstract
- Legionella is a pathogenic Gram-negative bacterium that can multiply inside of eukaryotic cells. It translocates numerous bacterial effector proteins into target cells to transform host phagocytes into a niche for replication. One effector of Legionella pneumophila is the glucosyltransferase Lgt1, which modifies serine 53 in mammalian elongation factor 1A (eEF1A), resulting in inhibition of protein synthesis and cell death. Here, we demonstrate that similar to mammalian cells, Lgt1 was severely toxic when produced in yeast and effectively inhibited in vitro protein synthesis. Saccharomyces cerevisiae strains, which were deleted of endogenous eEF1A but harbored a mutant eEF1A not glucosylated by Lgt1, were resistant toward the bacterial effector. In contrast, deletion of Hbs1, which is also an in vitro substrate of the glucosyltransferase, did not influence the toxic effects of Lgt1. Serial mutagenesis in yeast showed that Phe(54), Tyr(56) and Trp(58), located immediately downstream of serine 53 of eEF1A, are essential for the function of the elongation factor. Replacement of serine 53 by glutamic acid, mimicking phosphorylation, produced a non-functional eEF1A, which failed to support growth of S. cerevisiae. Our data indicate that Lgt1-induced lethal effect in yeast depends solely on eEF1A. The region of eEF1A encompassing serine 53 plays a critical role in functioning of the elongation factor.
- Subjects :
- Glycosylation
Saccharomyces cerevisiae Proteins
Legionella
Saccharomyces cerevisiae
Amino Acid Motifs
Biochemistry
Legionella pneumophila
Microbiology
Serine
Glucosyltransferases
Peptide Elongation Factor 1
Bacterial Proteins
GTP-Binding Proteins
HSP70 Heat-Shock Proteins
Molecular Biology
biology
Cell Biology
biology.organism_classification
bacterial infections and mycoses
Peptide Elongation Factors
Yeast
Peptide Fragments
Recombinant Proteins
respiratory tract diseases
Elongation factor
Phenotype
Amino Acid Substitution
Protein Biosynthesis
Host-Pathogen Interactions
biology.protein
Mutagenesis, Site-Directed
bacteria
Glucosyltransferase
Gene Deletion
Subjects
Details
- ISSN :
- 1083351X
- Volume :
- 287
- Issue :
- 31
- Database :
- OpenAIRE
- Journal :
- The Journal of biological chemistry
- Accession number :
- edsair.doi.dedup.....ac0f480c3d275a8b3aa490a8b942e19b