Extended Data Fig. 2: Analysis of growth and c-di-AMP levels in mutants and after antibiotic exposure.
From: Cyclic-di-AMP modulates cellular turgor in response to defects in bacterial cell wall synthesis
(A) Bar graph showing β-galactosidase activity from the riboswitch reporter of the indicated B. subtilis deletion mutants. Cells lacking cdaAR have lower c-di-AMP and cells lacking gdpP have higher c-di-AMP. All measurements were performed in biological triplicate except the ∆cdaS condition which was performed in duplicate. (B) Bar graph showing β-galactosidase activity from the kimA reporter in cells lacking disA, cdaS, and cdaAR, and harbouring an IPTG-regulated allele of cdaAR in the presence of 50 µM IPTG. The strain was treated with sub-inhibitory concentrations (0.5X MIC) of fosfomycin (FOS), bacitracin (BAC), penicillin G (PENG), oxacillin (OX), vancomycin (VAN), moenomycin (MOE), D-cycloserine (DCYC). Cells only increase c-di-AMP levels in the presence of moenomycin that inhibits the glycosyltransferase activity of class A PBPs. (C) Bar graph showing β-galactosidase activity from the kimA riboswitch reporter in cells expressing CdaAR or DisA as the only c-di-AMP cyclase in the presence of absence of PBP1. The center of error represents the average, and the error bars represent the standard deviation across three biological replicates. (D) 10-fold serial dilutions of the indicated strains spotted on LB agar supplemented with the indicated IPTG concentrations. CdaAR is critical in the absence of PBP1.
