Roles of GacSA and DJ41_1407 in Acinetobacter baumannii ATCC 19606.

Two-component systems (TCSs) in bacteria are often involved in the global regulation of various physiological activities and behaviours. This study investigated the GacSA TCS and DJ41_1407 transcriptional sensor adjacent to GacA in Acinetobacter baumannii ATCC 19606. The relationship between GacS, GacA, and DJ41_1407 and their functions and signal transduction mechanisms are described. A. baumannii ATCC 19606 mutants, ∆gacS, ∆gacA, and ∆DJ41_1407, were generated using markerless mutation and cultured in LB medium, then collected for RNA sequencing. It was found that GacS, GacA, and DJ41_1407 regulate a series of genes involved in carbon metabolism. Quantitative reverse transcription PCR (qRT-PCR) results showed that DJ41_1407 and GacA may regulate the expression of adh4, ipdC, iacH, and paa. Phos-tag™ results revealed that GacS plays a more significant role in GacA phosphorylation. GacA regulated colony size and growth conditions in rich medium. Compared to the wild-type strain, the ∆gacA and ∆gacSA mutants exhibited smaller colony sizes, and mutation of the gacS, gacA, and DJ41_1407 genes also reduced bacterial virulence as determined by the Galleria mellonella infection assay. GacA also plays a crucial role in modulating antibiotic resistance, and the ∆gacA∆DJ41_1407 mutant demonstrated greater susceptibility to antibiotics. These results highlight the multiple functions regulated by the GacSA global TCS in A. baumannii ATCC 19606.