Elongation Factor 1A Is the Target of Growth Inhibition in Yeast Caused by Legionella pneumophila Glucosyltransferase Lgt1

Belyi, Yury; Tartakovskaya, Dina; Tais, Arlette; Fitzke, Edith; Tzivelekidis, Tina; Jank, Thomas; Rospert, Sabine; Aktories, Klaus

doi:10.1074/jbc.m112.372672

Cited by 29 publications

(29 citation statements)

References 40 publications

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“…It should be noted that the stability of wild type protein is relatively less than the corresponding alanine variants. Similar observation has also been recorded earlier where mutants were found more stable than wild type [ 14 , 22 , 25 ].…”

Section: Resultssupporting

confidence: 90%

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Identification of Critical Amino Acids Conferring Lethality in VopK, a Type III Effector Protein of Vibrio cholerae: Lessons from Yeast Model System

et al. 2015

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VopK, a type III effector protein, has been implicated in the pathogenesis of Vibrio cholerae strains belonging to diverse serogroups. Ectopic expression of this protein exhibits strong toxicity in yeast model system. In order to map critical residues in VopK, we scanned the primary sequence guided by available data on various toxins and effector proteins. Our in silico analysis of VopK indicated the presence of predicted MCF1-SHE (SHxxxE) serine peptidase domain at the C-terminus region of the protein. Substitution of each of the predicted catalytic triad residues namely Ser314, His353 and Glu357 with alanine resulted in recombinant VopK proteins varying in lethality as evaluated in yeast model system. We observed that replacement of glutamate357 to alanine causes complete loss in toxicity while substitutions of serine314 and histidine353 with alanine exhibited partial loss in toxicity without affecting the stability of variants. In addition, replacement of another conserved serine residue at position 354 (S354) within predicted S314H353E357 did not affect toxicity of VopK. In essence, combined in silico and site directed mutagenesis, we have identified critical amino acids contributing to the lethal activity of VopK in yeast model system.

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Section: Resultssupporting

confidence: 90%

“…This can be explained in terms of leaky expression from GAL1 promoter of pESC-LEU vector. The leaky expression problem associated with GAL1/10 promoter from different vectors has also been documented in earlier studies [ 20 – 22 ].…”

Section: Resultsmentioning

confidence: 58%

Identification of Critical Amino Acids Conferring Lethality in VopK, a Type III Effector Protein of Vibrio cholerae: Lessons from Yeast Model System

et al. 2015

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“…[PSI ϩ ] and [psi Ϫ ] versions of the wildtype and ⌬zuo1 strains were previously described (44). Deletion strains lacking ZUO1 (⌬zuo1), SKI7 (⌬ski7), HBS1 (⌬hbs1), LTN1 (⌬ltn1), and the ⌬zuo1 ⌬ski7 and ⌬zuo1 ⌬ltn1 double deletion strains were described previously (32,34,51). Ssb is encoded by two closely related genes (SSB1 and SSB2), which are functionally interchangeable.…”

Section: Methodsmentioning

confidence: 99%

Release Factor eRF3 Mediates Premature Translation Termination on Polylysine-Stalled Ribosomes in Saccharomyces cerevisiae

Chiabudini

Tais

Zhang

et al. 2014

Molecular and Cellular Biology

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bRibosome stalling is an important incident enabling the cellular quality control machinery to detect aberrant mRNA. Saccharomyces cerevisiae Hbs1-Dom34 and Ski7 are homologs of the canonical release factor eRF3-eRF1, which recognize stalled ribosomes, promote ribosome release, and induce the decay of aberrant mRNA. Polyadenylated nonstop mRNA encodes aberrant proteins containing C-terminal polylysine segments which cause ribosome stalling due to electrostatic interaction with the ribosomal exit tunnel. Here we describe a novel mechanism, termed premature translation termination, which releases C-terminally truncated translation products from ribosomes stalled on polylysine segments. Premature termination during polylysine synthesis was abolished when ribosome stalling was prevented due to the absence of the ribosomal protein Asc1. In contrast, premature termination was enhanced, when the general rate of translation elongation was lowered. The unconventional termination event was independent of Hbs1-Dom34 and Ski7, but it was dependent on eRF3. Moreover, premature termination during polylysine synthesis was strongly increased in the absence of the ribosome-bound chaperones ribosome-associated complex (RAC) and Ssb (Ssb1 and Ssb2). On the basis of the data, we suggest a model in which eRF3-eRF1 can catalyze the release of nascent polypeptides even though the ribosomal A-site contains a sense codon when the rate of translation is abnormally low.

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“…The regulation of translation at the level of the interaction between eEF1A1 and eEF1B2—like we show here for TIP30—has been previously demonstrated as the result of eEF1B2 phosphorylation by the cell cycle‐dependent kinase (CDK)1 leading to reduced interaction with eEF1A1 and downregulation of translation during mitosis (Sivan et al , ; Sasikumar et al , ). Reduced activation of translation due to decreased eEF1A1 activity leads to diminished cell proliferation or cell growth in different cell types (Kim et al , ; Lin et al , ; Belyi et al , ). Remarkably, TIP30 appears to act on protein synthesis independent of mTORC1, which was even de‐activated in a counter‐regulatory manner in TIP30‐deficient mice.…”

Section: Discussionmentioning

confidence: 99%

TIP 30 counteracts cardiac hypertrophy and failure by inhibiting translational elongation

et al. 2019

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Pathological cardiac overload induces myocardial protein synthesis and hypertrophy, which predisposes to heart failure. To inhibit hypertrophy therapeutically, the identification of negative regulators of cardiomyocyte protein synthesis is needed. Here, we identified the tumor suppressor protein TIP30 as novel inhibitor of cardiac hypertrophy and dysfunction. Reduced TIP30 levels in mice entailed exaggerated cardiac growth during experimental pressure overload, which was associated with cardiomyocyte cellular hypertrophy, increased myocardial protein synthesis, reduced capillary density, and left ventricular dysfunction. Pharmacological inhibition of protein synthesis improved these defects. Our results are relevant for human disease, since we found diminished cardiac TIP30 levels in samples from patients suffering from end‐stage heart failure or hypertrophic cardiomyopathy. Importantly, therapeutic overexpression of TIP30 in mouse hearts inhibited cardiac hypertrophy and improved left ventricular function during pressure overload and in cardiomyopathic mdx mice. Mechanistically, we identified a previously unknown anti‐hypertrophic mechanism, whereby TIP30 binds the eukaryotic elongation factor 1A (eEF1A) to prevent the interaction with its essential co‐factor eEF1B2 and translational elongation. Therefore, TIP30 could be a therapeutic target to counteract cardiac hypertrophy.

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Elongation Factor 1A Is the Target of Growth Inhibition in Yeast Caused by Legionella pneumophila Glucosyltransferase Lgt1

Cited by 29 publications

References 40 publications

Identification of Critical Amino Acids Conferring Lethality in VopK, a Type III Effector Protein of Vibrio cholerae: Lessons from Yeast Model System

Identification of Critical Amino Acids Conferring Lethality in VopK, a Type III Effector Protein of Vibrio cholerae: Lessons from Yeast Model System

Release Factor eRF3 Mediates Premature Translation Termination on Polylysine-Stalled Ribosomes in Saccharomyces cerevisiae

TIP 30 counteracts cardiac hypertrophy and failure by inhibiting translational elongation

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