Antitoxin-induced auto-phosphorylation neutralizes the nucleotidyltransferase toxin AbiEii from Streptococcus agalactiae to safeguard global translation

Arrowsmith, T. J.; Xu, X.; Went, S. C.; Han, X.; Kelly, A.; Chansigaud, P.; Emsley, Z. J.; Dubrulle, J.; Fineran, P. C.; Genevaux, P.; Blower, T. R.

doi:10.64898/2026.01.19.700257

bioRxiv · 10.64898/2026.01.19.700257

Antitoxin-induced auto-phosphorylation neutralizes the nucleotidyltransferase toxin AbiEii from Streptococcus agalactiae to safeguard global translation

Arrowsmith, T. J.·Xu, X.·Went, S. C.·Han, X.·Kelly, A.·Chansigaud, P.·Emsley, Z. J.·Dubrulle, J.·Fineran, P. C.·Genevaux, P.·Blower, T. R.

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Abstract

Nucleotidyltransferases (NTases) control diverse physiological processes, including DNA replication and repair, antibiotic resistance, and RNA modification. The Streptococcus agalactiae abortive infection family toxin, AbiEii, belongs to the DUF1814 superfamily of NTases and blocks bacterial growth through an as yet unknown mechanism. Antitoxin AbiEi has been reported to antagonize AbiEii as part of a Type IV toxin-antitoxin system. Here, we demonstrate through structural, biochemical, and biophysical studies that AbiEi binds to AbiEii to form a stable toxin-antitoxin complex in solution. Antitoxin binding ultimately leads to AbiEii auto-phosphorylation and an abolition of NTase activity. These data reclassify AbiE systems as Type VII toxin-antitoxin systems. Toxin phosphorylation appears to be a dynamic and reversible process, and we show that the sole serine/threonine phosphatase SaSTP of S. agalactiae can efficiently dephosphorylate AbiEii in vitro. Mutagenesis studies and functional assays indicate that a widespread mechanism of toxin auto-phosphorylation accounts for AbiEii neutralization, further supporting recent hypotheses outlining control of homologous MenAT toxin-antitoxin systems from Mycobacterium tuberculosis.

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Arrowsmith, T. J., Xu, X., Went, S. C., Han, X., Kelly, A., Chansigaud, P., Emsley, Z. J., Dubrulle, J., Fineran, P. C., Genevaux, P., Blower, T. R.. 2026-01-20. Antitoxin-induced auto-phosphorylation neutralizes the nucleotidyltransferase toxin AbiEii from Streptococcus agalactiae to safeguard global translation. https://doi.org/10.64898/2026.01.19.700257

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