Thialysine utilization by E. coli and its effects on cell growth

Girolamo, M. Di; Busiello, V.; Cini, Chiara; Foppoli, Cesira; Marco, C. De

doi:10.1007/bf00215580

Cited by 11 publications

(7 citation statements)

References 7 publications

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“…In this way, the lysine analogue can be incorporated into polypeptide chains resulting in the formation of inactive proteins. Thialysine is toxic against bacteria, as well as against eukaryotic cells [24][25][26]. The data presented here indicate that, at least in the prokaryote E. coli, cytotoxicity of thialysine is prevented by N ε -acetylation mediated by the introduced C. elegans N-acetyltransferase D2023.4.…”

Section: Discussionmentioning

confidence: 76%

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A novel member of the GCN5-related N-acetyltransferase superfamily from Caenorhabditis elegans preferentially catalyses the N-acetylation of thialysine [S-(2-aminoethyl)-L-cysteine]

Abo-Dalo

Ndjonka

Pinnen

et al. 2004

Biochemical Journal

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The putative diamine N-acetyltransferase D2023.4 has been cloned from the model nematode Caenorhabditis elegans. The 483 bp open reading frame of the cDNA encodes a deduced polypeptide of 18.6 kDa. Accordingly, the recombinantly expressed His6-tagged protein forms an enzymically active homodimer with a molecular mass of approx. 44000 Da. The protein belongs to the GNAT (GCN5-related N-acetyltransferase) superfamily, and its amino acid sequence exhibits considerable similarity to mammalian spermidine/spermine-N1-acetyltransferases. However, neither the polyamines spermidine and spermine nor the diamines putrescine and cadaverine were efficiently acetylated by the protein. The smaller diamines diaminopropane and ethylenediamine, as well as L-lysine, represent better substrates, but, surprisingly, the enzyme most efficiently catalyses the N-acetylation of amino acids analogous with L-lysine. As determined by the k(cat)/K(m) values, the C. elegans N-acetyltransferase prefers thialysine [S-(2-aminoethyl)-L-cysteine], followed by O-(2-aminoethyl)-L-serine and S-(2-aminoethyl)-D,L-homocysteine. Reversed-phase HPLC and mass spectrometric analyses revealed that N-acetylation of L-lysine and L-thialysine occurs exclusively at the amino moiety of the side chain. Remarkably, heterologous expression of C. elegans N-acetyltransferase D2023.4 in Escherichia coli, which does not possess a homologous gene, results in a pronounced resistance against the anti-metabolite thialysine. Furthermore, C. elegans N-acetyltransferase D2023.4 exhibits the highest homology with a number of GNATs found in numerous genomes from bacteria to mammals that have not been biochemically characterized so far, suggesting a novel group of GNAT enzymes closely related to spermidine/spermine-N1-acetyltransferase, but with a distinct substrate specificity. Taken together, we propose to name the enzyme 'thialysine N(epsilon)-acetyltransferase'.

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Section: Discussionmentioning

confidence: 76%

“…Thialysine is a known potent anti-metabolite of L-lysine that inhibits growth of prokaryotic as well as eukaryotic cells [24][25][26].…”

Section: Heterologous Expression Of C Elegans N-acetyltransferase D2mentioning

confidence: 99%

A novel member of the GCN5-related N-acetyltransferase superfamily from Caenorhabditis elegans preferentially catalyses the N-acetylation of thialysine [S-(2-aminoethyl)-L-cysteine]

Abo-Dalo

Ndjonka

Pinnen

et al. 2004

Biochemical Journal

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“…One such compound of particular interest is the metabolic intermediate S-(2-aminoethyl)-Lcysteine (thialysine), a naturally occurring proteinogenic lysine analogue (15)(16)(17)(18)(19). Thialysine is a sufficiently good mimic of lysine that it can effectively prevent cellular growth by incorporation into protein via LysRS2 (20)(21)(22). The potential reduction in thialysine incorporation offered by the use of LysRS1 suggests that it might provide an effective means to restrict the analogue's detrimental effects.…”

mentioning

confidence: 99%

Nonorthologous replacement of lysyl-tRNA synthetase prevents addition of lysine analogues to the genetic code

Jester

Levengood

Roy

et al. 2003

Proc. Natl. Acad. Sci. U.S.A.

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Insertion of lysine during protein synthesis depends on the enzyme lysyl-tRNA synthetase (LysRS), which exists in two unrelated forms, LysRS1 and LysRS2. LysRS1 has been found in most archaea and some bacteria, and LysRS2 has been found in eukarya, most bacteria, and a few archaea, but the two proteins are almost never found together in a single organism. Comparison of structures of LysRS1 and LysRS2 complexed with lysine suggested significant differences in their potential to bind lysine analogues with backbone replacements. One such naturally occurring compound, the metabolic intermediate S-(2-aminoethyl)-L-cysteine, is a bactericidal agent incorporated during protein synthesis via LysRS2. In vitro tests showed that S-(2-aminoethyl)-L-cysteine is a poor substrate for LysRS1, and that it inhibits LysRS1 200-fold less effectively than it inhibits LysRS2. In vivo inhibition by S-(2-aminoethyl)-L-cysteine was investigated by replacing the endogenous LysRS2 of Bacillus subtilis with LysRS1 from the Lyme disease pathogen Borrelia burgdorferi. B. subtilis strains producing LysRS1 alone were relatively insensitive to growth inhibition by S-(2-aminoethyl)-L-cysteine, whereas a WT strain or merodiploid strains producing both LysRS1 and LysRS2 showed significant growth inhibition under the same conditions. These growth effects arising from differences in amino acid recognition could contribute to the distribution of LysRS1 and LysRS2 in different organisms. More broadly, these data demonstrate how diversity of the aminoacyltRNA synthetases prevents infiltration of the genetic code by noncanonical amino acids, thereby providing a natural reservoir of potential antibiotic resistance.

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“…[140] It was also established that AEC is incorporated into proteins, which could have toxic effects on the cell. [141,142] Various E. coli and B. subtilis mutant strains [143][144][145] resistant to AEC were found to cause derepression of lysC expression. The mutations were located in the lysine aptamer domain; [45] this suggests that the toxic effects might be due to the repression of aspartokinase expression following binding of AEC to lysine riboswitch.…”

Section: Riboswitches As Antibacterial Targetsmentioning

confidence: 99%

Riboswitches: Ancient and Promising Genetic Regulators

et al. 2009

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Bait and switch: Metabolite‐sensing riboswitches make use of RNA structural modulation to regulate gene expression, as illustrated in the scheme, in response to subtle changes in metabolite concentrations. This review describes the current knowledge about naturally occurring riboswitches and their growing potential as antibacterial cellular targets and as molecular biosensors.magnified imageNewly discovered metabolite‐sensing riboswitches have revealed that cellular processes extensively make use of RNA structural modulation to regulate gene expression in response to subtle changes in metabolite concentrations. Riboswitches are involved at various regulation levels of gene expression, such as transcription attenuation, translation initiation, mRNA splicing and mRNA processing. Riboswitches are found in the three kingdoms of life, and in various cases, are involved in the regulation of essential genes, which makes their regulation an essential part of cell survival. Because riboswitches operate without the assistance of accessory proteins, they are believed to be remnants of an ancient time, when gene regulation was strictly based on RNA, from which are left numerous “living molecular fossils”, as exemplified by ribozymes, and more spectacularly, by the ribosome. Due to their nature, riboswitches hold high expectations for the manipulation of gene expression and the detection of small metabolites, and also offer an unprecedented potential for the discovery of novel classes of antimicrobial agents.

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Thialysine utilization by E. coli and its effects on cell growth

Cited by 11 publications

References 7 publications

A novel member of the GCN5-related N-acetyltransferase superfamily from Caenorhabditis elegans preferentially catalyses the N-acetylation of thialysine [S-(2-aminoethyl)-L-cysteine]

A novel member of the GCN5-related N-acetyltransferase superfamily from Caenorhabditis elegans preferentially catalyses the N-acetylation of thialysine [S-(2-aminoethyl)-L-cysteine]

Nonorthologous replacement of lysyl-tRNA synthetase prevents addition of lysine analogues to the genetic code

Riboswitches: Ancient and Promising Genetic Regulators

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