Fig 3. V-59 stimulates Rv1625c to produce cAMP.
(A) Impact of V-59 on cAMP production in Mtb. Cultures were treated with V-59 or DMSO for 24 hours. Data are from two experiments with two technical replicates each (***P < 0.001, ****P < 0.0001, One-way ANOVA with Sidak’s multiple comparisons test). (B) Impact of Rv1625c agonists on cAMP production in cya- E. coli transformed with an empty vector control or an Rv1625c expression plasmid. Supernatants were collected 18 hours after addition of V-59, mCLB073, or DMSO. Data is from one experiment, with three independent expression clones, and two technical replicates each. In (A) and (B) Data are normalized as total cAMP per 108 bacteria. DMSO is the vehicle control. Data are shown as means ± SD. (C) Summary of mutations in the rv1625c gene that confer resistance to Rv1625c agonists. Mutations are grouped by their effect on the rv1625c sequence, with missense mutations (blue) and insertion or frameshift mutations (red) and mapped on the Rv1625c topology diagram to illustrate their approximate location relative to Rv1625c protein domains. Black circles represent amino acids that are essential for AC activity.