Individual Nudix hydrolases affect diverse features of Pseudomonas aeruginosa

Abstract

Nudix proteins catalyze the hydrolysis of pyrophosphate bonds in a variety of substrates and are ubiquitous in all domains of life. The genome of an important opportunistic human pathogen, Pseudomonas aeruginosa , encodes multiple Nudix proteins. To determine the role of nine Nudix hydrolases of the P. aeruginosa PAO1161 strain in its fitness, virulence or antibiotic resistance mutants devoid of individual enzymes were constructed and analyzed for growth rate, motility, biofilm formation, pyocyanin production, and susceptibility to oxidative stress and different antibiotics. The potential effect on bacterial virulence was studied using the C aenorhabditis elegans – P. aeruginosa infection model. Of the nine mutants tested, five had an altered phenotype in comparison with the wild‐type strain. The ΔPA3470, ΔPA3754, and ΔPA4400 mutants showed increased pyocyanin production, were more resistant to the β‐lactam antibiotic piperacillin, and were more sensitive to killing by H 2 O 2 . In addition, ΔPA4400 and ΔPA5176 had impaired swarming motility and were less virulent for C. elegans . The ΔPA4841 had an increased sensitivity to oxidative stress. These changes were reversed by providing the respective nudix gene in trans indicating that the observed phenotype alterations were indeed due to the lack of the particular Nudix protein.

Dual RNA-Seq of Flavobacterium psychrophilum and Its Outer Membrane Vesicles Distinguishes Genes Associated with Susceptibility to Bacterial Cold-Water Disease in Rainbow Trout (Oncorhynchus mykiss)

“…Furthermore, nudix proteins catalyze the hydrolysis of pyrophosphate bonds and have a demonstrated role in bacterial fitness and virulence. Mutants of Pseudomonas aeruginosa devoid of individual nudix hydrolases were more sensitive to killing by oxidative stress/H 2 O 2 and showed less virulence [ 52 ]. Moreover, Rhs proteins play a crucial role in the interaction between bacteria and host cell.…”

Functional Amyloids in Pseudomonas aeruginosa Are Essential for the Proteome Modulation That Leads to Pathoadaptation in Pulmonary Niches

“…There was significant downregulation of stringent starvation protein A (SspA) in the PAO1 Δfap SIP group; this protein provides adaptation to stringent stress responses in a new environment ( 39 ). A nudix pyrophosphatase (PA5176) relieved ROS generation and also contributed toward motility and pathogenicity ( 40 ). PA5176 was found to be upregulated ~8-fold in PAO1 pFap attached cells.…”

Pangenomic analysis reveals plant NAD+manipulation as an important virulence activity of bacterial pathogen effectors

“…Using HMMER-based approach, 146 candidate OGs were identified, whilst the HHsearch-based approach picked up 564 candidates, including 136 being identified by both approaches (Table 2, full details in Supplemental Table S2). Among the 136 candidates are known housekeeping nucleosidases, such as the NUDIX enzyme NudC (OG1452), the Sirtuin CobB3 (OG752), the purine nucleosidase RihA (OG2649), and cyclic-AMP phosphodiesterase CpdA (OG3903) (Drabinska et al, 2020; Fuchs et al, 2010; Gallego-Jara et al, 2021; Petersen and Moller, 2001). In addition, both searches identified the TIR T3E HopAM1 (OG6638) (Eastman et al, 2022) and the ART T3E HopU1 (OG9885) (Fu et al, 2007).…”

“…Although we focused on identifying T3Es in this study, it would be interesting to determine if any of these non-T3E NADases are hrp -regulated and thus may also contribute to pathogenesis. For example, Drabinska et al (2020) identified multiple NUDIX enzymes in the animal opportunistic pathogen P. aeruginosa that are involved in virulence, production of the toxin pyocyanin, and resistance to oxidative stress.…”