Global transcription changes driven by the second messenger cyclic di-GMP in Acinetobacter baumannii AB5075

In order to know the effect of the second messenger cyclic di-GMP (cdGMP) on the global transcription of A. baumannii AB5075, we performed a differential RNA sequencing (dRNA-seq) experiment comparing two AB5075 derivatives that allow IPTG-induced overexpression of the phosphodiesterase coding gene rocR (decreases cdGMP levels) or a constitutively active version of the diguanylate cyclase pleD (pleD*, increases cdGMP levels) A. baumannii AB5075 derivatives carrying either rocR or pleD* inserted in a miniTn7 transposon were grown in LB supplemented with IPTG 1 mM (3 biological replicates per strain) until reaching mid-exponential phase (approx.

OD600 0.6). At this point, cells were harvested, treated with RNAlater for preservation of total RNA and stored at -80 C. After that, total RNA was extracted from each sample. As a result, 154 genes appeared upregulated in high cdGMP conditions (overexpressing pleD*) whereas 188 were downregulated.