Several Kac modification sites of Aha1 affect OXY resistance and intracellular OXY accumulation.A–F, the antibiotic susceptibility assay of Δaha1 and site-directed mutagenesis generated aha1 mutants to OXY using the MBC plate method. The aha1 mutants are numbered in the following order: 1, WT; 2, Δaha1; 3, Δaha1+paha1; 4, Δaha1+vector; 5, Δaha1+paha1-K57R; 6, Δaha1+paha1-K57Q; 7, Δaha1+paha1-K99R; 8, Δaha1+paha1-K99Q; 9, Δaha1+paha1-K166R; 10, Δaha1+paha1-K166Q; 11, Δaha1+paha1-K175R; 12, Δaha1+paha1-K175Q; 13, Δaha1+paha1-K178R; 14, Δaha1+paha1-K178Q; 15, Δaha1+paha1-K187R; 16, Δaha1+paha1-K187Q; 17, Δaha1+paha1-K197R; 18, Δaha1+paha1-K197Q; 19, Δaha1+paha1-K57R-K187Q-K197Q; 20, Δaha1+paha1-K57Q-K187R-K197Q; 21, Δaha1+paha1-K57Q-K187R-K197R; 22, Δaha1+paha1-K57R-K187R-K197R; 23, Δaha1+paha1-K57Q-K187Q-K197R; 24, Δaha1+paha1-K57Q-K187Q-K197Q; 25, Δaha1+paha1-K57R-K187R-K197Q; 26, Δaha1+paha1-K57R-K187Q-K197R. G, determination of intracellular OXY accumulations in aha1 mutants under 2 mM OXY for 2 h. OXY, oxytetracycline; Kac, lysine acetylation; MBC, minimal bacterial concentration.