The role and relationship with efflux pump of biofilm formation in Klebsiella pneumoniae

Tang, Miran; Wang, Xing; Wan, Xue; Ding, Zixuan; Ding, Yinhuan; Wang, Zhibin

doi:10.1016/j.micpath.2020.104244

Cited by 43 publications

(23 citation statements)

References 38 publications

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“…As one of the largest superfamily of secondary carriers known to date, MFS family membrane transporters exist widely in the whole biological world ( Reddy et al, 2012 ) and they are considered to be closely related to a wide variety of life phenomena due to a basic function of assisting in transmembrane transport of certain substances, including monosaccharides, oligosaccharides, amino acids, enzyme cofactors, drugs, and so on ( Lorca et al, 2007 ; Chen et al, 2008 ; Yen et al, 2010 ). In bacteria, MFS superfamily proteins have been found not only to play an important role in the transport of many substances but also to be a member of the efflux pumps large families ( Pasqua et al, 2019 ), which are involved in biofilm formation and drug resistance ( Bay et al, 2017 ; Saranathan et al, 2017 ; Poudyal and Sauer, 2018 ; Tang et al, 2020 ).…”

Section: Discussionmentioning

confidence: 99%

Overexpression of BIT33_RS14560 Enhances the Biofilm Formation and Virulence of Acinetobacter baumannii

Yang

Lai²,

Liao³

et al. 2022

Front. Microbiol.

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Acinetobacter baumannii, a strictly aerobic, non-lactose fermented Gram-negative bacteria, is one of the important pathogens of nosocomial infection. Major facilitator superfamily (MFS) transporter membrane proteins are a class of proteins that widely exists in microbial genomes and have been revealed to be related to biofilm formation in a variety of microorganisms. However, as one of the MFS transporter membrane proteins, little is known about the role of BIT33_RS14560 in A. baumannii. To explore the effects of BIT33_RS14560 on biofilm formation of A. baumannii, the biofilm formation abilities of 62 isolates were firstly investigated and compared with their transcript levels of BIT33_RS14560. Then, this specific gene was over-expressed in a standard A. baumannii strain (ATCC 19606) and two isolates of extensively drug-resistant A. baumannii (XDR-Ab). Bacterial virulence was observed using a Galleria mellonella infection model. High-throughput transcriptome sequencing (RNA seq) was performed on ATCC 19606 over-expressed strain and its corresponding empty plasmid control strain. Spearman’s correlation analysis indicated a significant negative correlation (R = −0.569, p = 0.000) between the △CT levels of BIT33_RS1456 and biofilm grading of A. baumannii isolates. The amount of A. baumannii biofilm was relatively high within 12–48 h. Regardless of standard or clinical strains; the biofilm biomass in the BIT33_RS14560 overexpression group was significantly higher than that in the control group ( p < 0.0001). Kaplan–Meier survival curve analysis showed that the mortality of G. mellonella was significantly higher when infected with the BIT33_RS14560 overexpression strain (χ2 = 8.462, p = 0.004). RNA-Seq showed that the mRNA expression levels of three genes annotated as OprD family outer membrane porin, glycosyltransferase family 39 protein, and glycosyltransferase family 2 protein, which were related to bacterial adhesion, biofilm formation, and virulence, were significantly upregulated when BIT33_RS14560 was over-expressed. Our findings provided new insights in identifying potential drug targets for the inhibition of biofilm formation. We also developed a practical method to construct an over-expressed vector that can stably replicate in XDR-Ab isolates.

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Section: Discussionmentioning

confidence: 99%

Overexpression of BIT33_RS14560 Enhances the Biofilm Formation and Virulence of Acinetobacter baumannii

Yang

Lai²,

Liao³

et al. 2022

Front. Microbiol.

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“…There is a growing body of evidence suggesting multiple role of the drug efflux pumps in biofilm formation like extrusion of extracellular polymeric substances, quorum sensing, and genetic regulation and facilitating the aggregation of the cells to establish biofilm matrix formation (Alav et al, 2018). Of late, biofilms were reported to have strong correlation with increased expression of efflux pump genes, namely, AcrA , emr B, oqx A, and qac EΔ1 in Klebsiella pneumoniae (Tang et al, 2020). In this study too, we observed a significantly elevated relative expression of AcrA among the strong biofilm producers compared with that among the weak producers.…”

Section: Discussionmentioning

confidence: 99%

Elucidating the resistance repertoire, biofilm production, and phylogenetic characteristics of multidrug‐resistant Escherichia coli isolated from community ponds: A study from West Bengal, India

Bhattacharyya

Banerjee

Habib

et al. 2022

Water Environment Research

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This study details about the phenotypic and molecular characteristics of multidrug‐resistant (MDR) Escherichia coli in the fresh community pond water (n = 257) collected from three districts of West Bengal, India. In total, 57 isolates were MDR of which 38 emerged as extended spectrum and 7 as AmpC‐type β‐lactamase producers in phenotypic assay. Among β‐lactamase genes, blaCTXM‐1was predominant (87.71%) followed by blaAmpC (77.2%) and blaTEM‐1 (22.8%). Six MDR strains carried metallo‐β‐lactamase (MBL, blaNDM‐1) gene. Tissue culture plate assay confirmed strong biofilm (SP) production in four MDR and one non‐MDR isolates. In PCR‐based replicon typing (PBRT), multiple plasmids of diverse replicon types (Frep, FIB, I1, FIA, K/B, HI1, and Y) were identified. The enterobacterial repetitive intergenic consensus–polymerase chain reaction (ERIC‐PCR)‐based phylogenetic analysis revealed a high degree of genetic divergence among the MDR isolates. Multiplex PCR‐based phylogrouping categorized 11 isolates as virulent (B2/D/F), which carried blaCTXM‐1 gene and three had blaNDM‐1 gene. Relative transcriptional activity of AcrAB efflux pump was significantly elevated among the SP and MBL producers. The presence of MDR E. coli isolates, particularly those resistant to carbapenem, in pond water used for daily domestic and household work, is a cause of concern as these pathogens may sneak into human food chain causing life‐threatening infections. Practitioner Points Multidrug‐resistant biofilm producing E. coli isolated from community pond water. A few of them were carbapenem‐resistant and belonged to virulent (B2/D) types. Expression of AcrAB efflux pumps was found significantly elevated among biofilm producers and carbapenem‐resistant population.

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“…As previously mentioned, biofilms of the wild‐type (WT) strain, ykgK mutant, and complementary strains and RcsAB mutant and complementary strains were identified [30,31]. In short, the cells were cultured in a glass tube of appropriate volume and incubated at 37°C for 36, 48, and 60 h. After the culture broth was transferred to a 96‐well microtiter plate, the OD600 value was measured with a plate reader (Varioskan LUX; Thermo Fisher Scientific).…”

Section: Methodsmentioning

confidence: 99%

Regulation of ECP fimbriae‐related genes by the transcriptional regulator RcsAB in Klebsiella pneumoniae NTUH‐K2044

Shen

Zhang²,

Xue

et al. 2022

J Basic Microbiol

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Klebsiella pneumoniae is one of the major pathogens causing nosocomial infections.The regulator of capsule synthesis (Rcs) system is a complex signal transduction pathway that is involved in the regulation of virulence factors of K. pneumoniae as an important transcriptional regulator. The RcsAB box-like sequence was found to be present in the promoter-proximal regions of ykgK, one of the ECP fimbriaerelated genes, which suggested the expression of ECP fimbriae may be regulated by RcsAB. The ykgK gene in K. pneumoniae has 86% similarity to the ecpR gene in Escherichia coli. Nucleotide sequence alignment revealed a similar ECP fimbriae gene cluster including six genes in K. pneumoniae, which was proved to be on the same operon in this study. The electrophoretic mobility shift assay and DNase I assay, relative fluorescence expression, β-galactosidase activity, and relative gene expression of ykgK in the wild-type and mutant strains were performed to determine the transcriptional regulation mechanism of RcsAB on ECP fimbriae. The mutant ΔykgK and complementary strain ΔykgK/cΔykgK were constructed to complete the Galleria mellonella larvae infection experiment and biofilm formation assay. This study showed that RcsAB binds directly to the promoter region of the ykgK gene to positively regulate ECP fimbriae-related gene clusters, and then positively affect the biofilm formation. K E Y W O R D S ECP fimbriae-related gene, Klebsiella pneumoniae, RcsAB, transcriptional regulator, ykgK gene 1 | INTRODUCTION Klebsiella pneumoniae is a common opportunistic pathogen that can cause urinary and respiratory tract infections, pneumonia, bacteraemia, and sepsis [1]. Current research on the virulence of K. pneumoniae has also attracted much attention. The known virulence factors of K. pneumoniae mainly include capsular polysaccharide (CPS), lipopolysaccharide, fimbriae, urease, biofilm formation-related factors, and the iron capture

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The role and relationship with efflux pump of biofilm formation in Klebsiella pneumoniae

Cited by 43 publications

References 38 publications

Overexpression of BIT33_RS14560 Enhances the Biofilm Formation and Virulence of Acinetobacter baumannii

Overexpression of BIT33_RS14560 Enhances the Biofilm Formation and Virulence of Acinetobacter baumannii

Elucidating the resistance repertoire, biofilm production, and phylogenetic characteristics of multidrug‐resistant Escherichia coli isolated from community ponds: A study from West Bengal, India

Regulation of ECP fimbriae‐related genes by the transcriptional regulator RcsAB in Klebsiella pneumoniae NTUH‐K2044

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