GEK1, a gene product of Arabidopsis thaliana involved in ethanol tolerance, is a D-aminoacyl-tRNA deacylase

Wydau, Sandra; Ferri-Fioni, Maria-Laura; Blanquet, Sylvain; Plateau, Pierre

doi:10.1093/nar/gkl1145

Cited by 27 publications

(34 citation statements)

References 32 publications

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“…Total amounts of proteins in all extracts were measured by using the Bradford protein assay (Bio-Rad), with bovine serum albumin as standard. 14 C]Lys-tRNA Lys (310 Ci/mol) were prepared from E. coli tRNAs as described previously (6,11,14,16).…”

Section: Preparation Of Crude Extracts-d-tyr-trnamentioning

confidence: 99%

“…Measurement of Generation Times-E. coli generation times were measured as described previously (16). To start the growth of cyanobacteria, Synechocystis strains PCC6803 and PCC6803⌬DTD3 were inoculated in 20 ml of the medium under study.…”

Section: Measurement Of Deacylation Rates-d-[mentioning

confidence: 99%

“…In minimal medium, the generation time of the E. coli ⌬dtd strain K37⌬RecA⌬TyrHDE3 is significantly increased (1.5-fold) upon addition of millimolar amounts of D-tyrosine (16). Thus, this strain was used to examine whether introduction of DTD3 could confer E. coli resistance against D-tyrosine.…”

Section: Expression Of Synechocystis Dtd3 Cures E Coli For Inactivatmentioning

confidence: 99%

“…In Arabidopsis thaliana, gek1 confers ethanol resistance. The product of this gene, surprisingly, behaves as a DTD2 deacylase (16). Consultation of the Pfam site (in August 2008) revealed that in a few bacteria (Aquifex aeolicus, Magnetococcus sp.…”

Section: D-aminoacyl-trna Deacylase Of Cyanobacteriamentioning

confidence: 99%

“…Importing dtd2 into E. coli functionally compensates for dtd deprivation. As shown in Table 1, the DTD2 protein has homologs in most archaea and in plants (16).…”

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Widespread Distribution of Cell Defense against d-Aminoacyl-tRNAs

Wydau¹,

Rest

Aubard³

et al. 2009

Journal of Biological Chemistry

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Several L-aminoacyl-tRNA synthetases can transfer a D-amino acid onto their cognate tRNA(s). This harmful reaction is counteracted by the enzyme D-aminoacyl-tRNA deacylase. Two distinct deacylases were already identified in bacteria (DTD1) and in archaea (DTD2), respectively. Evidence was given that DTD1 homologs also exist in nearly all eukaryotes, whereas DTD2 homologs occur in plants. On the other hand, several bacteria, including most cyanobacteria, lack genes encoding a DTD1 homolog. Here we show that Synechocystis sp. PCC6803 produces a third type of deacylase (DTD3). Inactivation of the corresponding gene (dtd3) renders the growth of Synechocystis sp. hypersensitive to the presence of D-tyrosine. Based on the available genomes, DTD3-like proteins are predicted to occur in all cyanobacteria. Moreover, one or several dtd3-like genes can be recognized in all cellular types, arguing in favor of the nearubiquity of an enzymatic function involved in the defense of translational systems against invasion by D-amino acids.Although they are detected in various living organisms (reviewed in Ref. 1), D-amino acids are thought not to be incorporated into proteins, because of the stereospecificity of aminoacyl-tRNA synthetases and of the translational machinery, including EF-Tu and the ribosome (2). However, the discrimination between L-and D-amino acids by aminoacyl-tRNA synthetases is not equal to 100%. Significant D-aminoacylation of their cognate tRNAs by Escherichia coli tyrosyl-, tryptophanyl-, aspartyl-, lysyl-, and histidyl-tRNA synthetases has been characterized in vitro (3-9). Recently, using a bacterium, transfer of D-tyrosine onto tRNA Tyr was shown to occur in vivo (10). With such misacylation reactions, the resulting D-aminoacyl-tRNAs form a pool of metabolically inactive molecules, at best. At worst, D-aminoacylated tRNAs infiltrate the protein synthesis machinery. Although the latter harmful possibility has not yet been firmly established, several cells were shown to possess a D-tyrosyl-tRNA deacylase, or DTD, that should help them counteract the accumulation of D-aminoacyl-tRNAs. This enzyme shows a broad specificity, being able to remove various D-aminoacyl moieties from the 3Ј-end of a tRNA (4 -6, 11). Such a function makes the deacylase a member of the family of enzymes capable of editing in trans mis-aminoacylated tRNAs. This family includes several homologs of aminoacyl-tRNA synthetase editing domains (12), as well as peptidyl-tRNA hydrolase (13,14).Two distinct deacylases have already been discovered. The first one, called DTD1, is predicted to occur in most bacteria and eukaryotes (see Table 1). Inactivation of the gene of this deacylase in E. coli (dtd) or in Saccharomyces cerevisiae (DTD1) exacerbates cell growth inhibition by several D-amino acids, including D-tyrosine (6). In fact, in an E. coli ⌬dtd strain grown in the presence of 2.4 mM D-tyrosine, as much as 40% of the cellular tRNATyr pool becomes esterified with D-tyrosine (10). Homologs of dtd/DTD1 are not found in the available archae...

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Section: Preparation Of Crude Extracts-d-tyr-trnamentioning

confidence: 99%

Section: Measurement Of Deacylation Rates-d-[mentioning

confidence: 99%

Section: Expression Of Synechocystis Dtd3 Cures E Coli For Inactivatmentioning

confidence: 99%

Section: D-aminoacyl-trna Deacylase Of Cyanobacteriamentioning

confidence: 99%

“…Importing dtd2 into E. coli functionally compensates for dtd deprivation. As shown in Table 1, the DTD2 protein has homologs in most archaea and in plants (16).…”

mentioning

confidence: 99%

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Widespread Distribution of Cell Defense against d-Aminoacyl-tRNAs

Wydau¹,

Rest

Aubard³

et al. 2009

Journal of Biological Chemistry

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Chiral proofreading during protein biosynthesis and its evolutionary implications

2022

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Homochirality of biomacromolecules is a prerequisite for their proper functioning and hence essential for all life forms. This underscores the role of cellular chiral checkpoints in enforcing homochirality during protein biosynthesis. d‐Aminoacyl‐tRNA deacylase (DTD) is an enzyme that performs ‘chirality‐based proofreading’ to remove d‐amino acids mistakenly attached to tRNAs, thus recycling them for further rounds of translation. Paradoxically, owing to its l‐chiral rejection mode of action, DTD can remove glycine as well, which is an achiral amino acid. However, this activity is modulated by discriminator base (N73) in tRNA, a unique element that protects the cognate Gly‐tRNAGly. Here, we review our recent work showing various aspects of DTD and tRNAGly coevolution and its key role in maintaining proper translation surveillance in both bacteria and eukaryotes. Moreover, we also discuss two major optimization events on DTD and tRNA that resolved compatibility issues among the archaeal and the bacterial translation apparatuses. Importantly, such optimizations are necessary for the emergence of mitochondria and successful eukaryogenesis.

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