Absence of tmRNA Increases the Persistence to Cefotaxime and the Intercellular Accumulation of Metabolite GlcNAc in Aeromonas veronii

Abstract

Bacterial persisters are a small proportion of phenotypically heterogeneous variants with the transient capability to survive in high concentrations of antibiotics, causing recurrent infections in both human and aquatic animals. Transfer-messenger RNA (tmRNA), which was encoded by the ssrA gene, was identified as a determinant regulator mediating the persistence to β-lactams in the pathogenic Aeromonas veronii C4. The deletion of tmRNA exhibited the increased ability of persister formation most probably due to the reduction of protein synthesis. Transcriptomic and metabolomic analyses revealed that the absence of tmRNA not only significantly elevated the intercellular levels of metabolite GlcNAc and promoted NaCl osmotic tolerance, but also upregulated the expression of metabolic genes in both the upstream biosynthesis pathway and the downstream metabolic flux of peptidoglycan (PG) biosynthesis. Finally, exogenous GlcNAc stimulated significant bacterial growth, enhanced content of GlcNAc in the cell wall, higher resistance to osmotic response, and higher persistence to cefotaxime in a concentration-dependent manner, implying its potential role in promoting the multiple phenotypes observed in tmRNA deletion strains. Taken together, these results hint at a potential mechanism of persister formation mediated by tmRNA against the β-lactam challenges in A. veronii.

Escherichia coli cryptic prophages sense nutrients to influence persister cell resuscitation

“…To determine whether cryptic prophages affect persister cell formation, we converted the wild‐type, the Δ9 strain with all nine cryptic prophages deleted (Wang et al ., 2010), and each single cryptic prophage mutant (Wang et al ., 2010) to persister cells using a rifampicin pretreatment (Kwan et al ., 2013) and enumerated them. This method of generating persister cells has been validated eight ways (Kim et al ., 2018b) and utilized by at least 15 independent labs to date (Johnson and Levin, 2013; Kwan et al ., 2013; Grassi et al ., 2017; Cui et al ., 2018; Jin et al ., 2018; Narayanaswamy et al ., 2018; Sulaiman et al ., 2018; Tkhilaishvili et al ., 2018; Pu et al ., 2019; Martins et al ., 2020; Rowe et al ., 2020; Sun et al ., 2020; Yu et al ., 2020; Zhao et al ., 2020; Zheng et al ., 2020); the approach has the benefit of increasing the number of persister cells by 10 000‐fold (Kwan et al ., 2013) which enables single‐cell studies (Kim et al ., 2018a; Kim et al ., 2018b; Song and Wood, 2020a; Yamasaki et al ., 2020).…”

Vibrio splendidus persister cells induced by host coelomic fluids show a similar phenotype to antibiotic‐induced counterparts

“…Whether the coelomic fluids‐induced and antibiotic‐resistant cells are bona fide persister cells is an important question in our study. In our experiments, we used several experimental methods to characterize persister cells (Kim et al ., 2018), which have also been validated by 15 independent labs to date (Johnson and Levin, 2013; Kwan et al ., 2013; Grassi et al ., 2017; Cui et al ., 2018; Jin et al ., 2018; Narayanaswamy et al ., 2018; Sulaiman et al ., 2018; Tkhilaishvili et al ., 2018; Pu et al ., 2019; Martins et al ., 2020; Rowe et al ., 2020; Sun et al ., 2020; Yu et al ., 2020; Zhao et al ., 2020; Zheng et al ., 2020). These methods are mainly based on the phenotypes of low metabolic activity, high gene expression of ppGpp‐pathway related genes, invariable MIC, multidrug tolerance, pathogenicity of re‐grown persister cells, and high phenotype heterogeneity.…”

Escherichia coliCryptic Prophages Sense Nutrients to Control Persister Cell Resuscitation

“…To determine whether cryptic prophages affect persister cell formation, we converted the wild-type, the Δ9 strain with all nine cryptic prophages deleted 2 , and each single cryptic prophage mutant 2 to persister cells using a rifampicin pretreatment 9 and enumerated them. This method of generating persister cells has been validated eight ways 19 and utilized by at least 15 independent labs to date 9,20,[23][24][25][26][27][28][29][30][31][32][33][34][35] ; the approach has the benefit of increasing the number of persister cells by 10,000-fold 9 which enables single-cell studies 3,5,15,19 . We found the presence of the cryptic prophages had no significant impact on the number of persister cells formed with ampicillin (Fig.…”