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. 2023 Dec 20;52(3):1298–1312. doi: 10.1093/nar/gkad1207

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© The Author(s) 2023. Published by Oxford University Press on behalf of Nucleic Acids Research.

This is an Open Access article distributed under the terms of the Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0/), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited.

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Figure 3. — Mutations in TifA that strengthen its interaction with ToxN. (A) Spotting assays for evolved T4 phage with increased expression of dmd-tifA(F41S) (T4 evo 2) and Thu1/R2 phage on lawns of +toxIN and +toxI-toxN(K111R) cells. (B) Alignment of residues 35–50 of TifA homologs from several T4-like phages, with residue 41 indicated. Degree of sequence similarity is highlighted in gray/black. (C) AlphaFold model for the ToxN-TifA_T4 complex, with residue F41 on TifA highlighted. (D) Serial dilutions of E. coli cells ectopically expressing toxN and wild-type or mutant tifA_T4 or tifA_RB69. (E) Western blot of ToxN-His₆ in cell lysate and following co-immunoprecipitation with TifA_T4-FLAG (wild-type or mutant) during T4 infection.