Fig. 1: DinB1Mtb activity requires five N-terminal amino acids omitted from the annotated ORF.
a, d, g Growth and b, f viability of strains carrying an inducible (tet = Anhydrotetracycline inducible promoter) DinB1 or its indicated derivatives (Msm = M. smegmatis, Mtb = annotated M. tuberculosis DinB1, Mtb+5aa = N terminal extended DinB1, dinB1 D113A = catalytically inactive M. smegmatis DinB1, Δβ clamp (ΔQESLF: 356–360 amino acids of M. smegmatis DinB1)) in presence of inducer. For a, the plotted OD value is the result of continuous dilution to maintain log phase growth (see methods). The viability in f was measured 24 h after inducer addition. c Anti-RecA/RpoB immunoblot from indicated strains with indicated times of inducer treatment. e Alignment of Msm and Mtb DinB1 N-termini with the potential start codons underlined. The blue boxed valine corresponds to the start codon of the published DinB1 noted as DinB1Mtb above, whereas the red boxed valine shows an alternative start codon of an extended DinB1 denoted as DinB1Mtb+5aa. Results shown are means (±SEM) of biological triplicates (a, b, d, g) or from biological replicates symbolized by gray dots (f). Stars above or under the means mark a statistical difference with the reference strain (empty vector) and lines connecting two strains show a statistical difference between them (*, P < 0.05; **, P < 0.01; ***, P < 0.001). p values were obtained on log-transformed data by one-way (f) or two-way (a, b, d, and g) ANOVA with a Bonferroni post-test.
