Characterization of a membrane toxin-antitoxin system, tsaAT, from Staphylococcus aureus.

Publisher: Published by Blackwell Pub. on behalf of the Federation of European Biochemical Societies Country of Publication: England NLM ID: 101229646 Publication Model: Print-Electronic Cited Medium: Internet ISSN: 1742-4658 (Electronic) Linking ISSN: 1742464X NLM ISO Abbreviation: FEBS J Subsets: MEDLINE

Original Publication: Oxford, UK : Published by Blackwell Pub. on behalf of the Federation of European Biochemical Societies, c2005-

Staphylococcus aureus*/genetics ; Staphylococcus aureus*/metabolism ; Toxin-Antitoxin Systems*/genetics ; Bacterial Toxins*/genetics ; Bacterial Toxins*/metabolism ; Cell Membrane*/metabolism ; Bacterial Proteins*/genetics ; Bacterial Proteins*/metabolism; Membrane Proteins/genetics ; Membrane Proteins/metabolism ; Amino Acid Sequence ; Amino Acid Substitution

Bacterial toxin-antitoxin (TA) systems consist of a toxin that inhibits essential cellular processes, such as DNA replication, transcription, translation, or ATP synthesis, and an antitoxin neutralizing their cognate toxin. These systems have roles in programmed cell death, defense against phage, and the formation of persister cells. Here, we characterized the previously identified Staphylococcus aureus TA system, tsaAT, which consists of two putative membrane proteins: TsaT and TsaA. Expression of the TsaT toxin caused cell death and disrupted membrane integrity, whereas TsaA did not show any toxicity and neutralized the toxicity of TsaT. Furthermore, subcellular fractionation analysis demonstrated that both TsaA and TsaT localized to the cytoplasmic membrane of S. aureus expressing either or both 3xFLAG-tagged TsaA and 3xFLAG-tagged TsaT. Taken together, these results demonstrate that the TsaAT TA system consists of two membrane proteins, TsaA and TsaT, where TsaT disrupts membrane integrity, ultimately leading to cell death. Although sequence analyses showed that the tsaA and tsaT genes were conserved among Staphylococcus species, amino acid substitutions between TsaT orthologs highlighted the critical role of the 6th residue for its toxicity. Further amino acid substitutions indicated that the glutamic acid residue at position 63 in the TsaA antitoxin and the cluster of five lysine residues in the TsaT toxin are involved in TsaA's neutralization reaction. This study is the first to describe a bacterial TA system wherein both toxin and antitoxin are membrane proteins. These findings contribute to our understanding of S. aureus TA systems and, more generally, give new insight into highly diverse bacterial TA systems.
(© 2024 The Author(s). The FEBS Journal published by John Wiley & Sons Ltd on behalf of Federation of European Biochemical Societies.)

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JP19KK0409 Japan Society for the Promotion of Science; JP20K07480 Japan Society for the Promotion of Science

Keywords: Staphylococcus aureus; membrane protein; toxin‐antitoxin system

0 (Bacterial Toxins)
0 (Bacterial Proteins)
0 (Membrane Proteins)

Date Created: 20241002 Date Completed: 20241113 Latest Revision: 20241113

20241114

10.1111/febs.17289

39356479