The LysR-Type Transcriptional Regulator YeeY Plays Important Roles in the Regulatory of Furazolidone Resistance in Aeromonas hydrophila

Abstract

Aeromonas hydrophila is an aquatic pathogen of freshwater fish. The emergence of widespread antimicrobial-resistance strains of this pathogen has caused increasing rates of fish infections. Our previous research reported that A. hydrophila yeeY, a LysRtype transcriptional regulator (LTTR), negatively regulated furazolidone (FZ) resistance. Although, it's intrinsic regulatory mechanism is still unclear. In this study, a dataindependent acquisition (DIA) quantitative proteomics method was used to compare the differentially expressed proteins (DEPs) between the yeeY and wild-type strain under FZ treatment. When compared to the control, a total of 594 DEPs were identified in yeeY. Among which, 293 and 301 proteins were substantially increased and decreased in abundance, respectively. Bioinformatics analysis showed that several biological pathways such as the secretion system and protein transport were mainly involved in FZ resistance. Subsequently, the antibiotics susceptibility assays of several gene deletion strains identified from the proteomics results showed that YeeY may regulate some important genes such as cysD, AHA_2766, AHA_3195, and AHA_4275, which affects the FZ resistance in A. hydrophila. Furthermore, 34 antimicrobial resistance genes (ARGs) from the bacterial drug resistance gene database (CARD) were found to be directly or indirectly regulated by YeeY. A subsequent assay of several ARGs mutants showed that AHA_3222 increased the susceptibility of A. hydrophila to FZ, while cysN and AHA_3753 decreased the susceptibility rate. Finally, the chromatin immunoprecipitation (ChIP) PCR and an electrophoretic mobility shift assay (EMSA) have revealed that the genes such as AHA_3222 and AHA_4275 were directly and transcriptionally regulated by YeeY. Taken together, our findings demonstrated that YeeY may participate in antimicrobial resistance of A. hydrophila to FZ, which provides a new target for the development of novel antimicrobial agents in the future.

Genomic Islands Identified in Highly Resistant Serratia sp. HRI: A Pathway to Discover New Disinfectant Resistance Elements

“…LysR-type transcriptional regulators have been reported to play a role in antibiotic resistance in Aeromonas sp. [ 43 ]. LuxR transcriptional regulators are involved in biofilm formation and stress response in Pseudomonas and Mycobacterium sp.…”

Tobramycin Stress Induced Differential Gene Expression in Acinetobacter baumannii

“…Upregulation of cysD has been shown in M. tuberculosis in response to nutritional deprivation, hypoxia, hydrogen peroxide, and vancomycin treatment [32]. cysD deletion mutant of Aeromonas hydrophila showed decreased survival rate against furazolidone treatment [33]. Since aminoglycosides produce reactive oxygen species [34], the downregulation of tauD and cysD genes in A. baumannii might have compromised the management of oxidative stress related to tobramycin.…”

Unveiling a Virulence-Regulating Mechanism in Aeromonas hydrophila: a Quantitative Exoproteomic Analysis of an AraC-Like Protein

“…The interaction between transcription factor orf02889 and candidate genes was verified by ChIP-PCR as previously described ( 23 ). Briefly, Δorf02889-C and Δorf02889-EV strains were transferred to 50 ml of LB medium, incubated at 200 rpm overnight, and then centrifuged at 5,000 g at 4°C for 10 min.…”