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LasB activation in Pseudomonas aeruginosa: Quorum sensing-mediated release of an auto-activation inhibitor
Cheol Seung Lee, Xi-Hui Li, Chae-Ran Jeon, Joon-Hee Lee
J. Microbiol. 2025;63(2):e2411005.   Published online February 27, 2025
DOI: https://doi.org/10.71150/jm.2411005
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  • 1 Scopus
AbstractAbstract PDF

Pseudomonas aeruginosa secretes three major proteases: elastase B (LasB), protease IV (PIV), and elastase A (LasA), which play crucial roles in infection and pathogenesis. These proteases are activated sequentially from LasB in a proteolytic cascade, and LasB was previously thought to undergo auto-activation. However, our previous study suggested that LasB cannot auto-activate independently but requires additional quorum sensing (QS)-dependent factors for activation, as LasB remained inactive in QS-deficient P. aeruginosa (QS-) even under artificial overexpression. In this study, we provide evidence for the existence of a LasB inhibitor in QS- mutants: inactive LasB overexpressed in QS- strains was in its processed form and could be reactivated upon purification; when full-length LasB was overexpressed in Escherichia coli, a heterologous bacterium lacking both LasB activators and inhibitors, the protein underwent normal processing and activation; and purified active LasB was significantly inhibited by culture supernatant (CS) from QS- strains but not by CS from QS+ strains. These findings demonstrate that a LasB inhibitor exists in QS- strains, and in its absence, LasB can undergo auto-activation without requiring an activator. Based on these results, we propose an updated hypothesis: the QS-dependent LasB activator functions by removing the LasB inhibitor rather than acting directly on LasB itself, thus preventing premature LasB activation until QS response is initiated.

Journal Articles
Application of fast expectation-maximization microbial source tracking to discern fecal contamination in rivers exposed to low fecal inputs
Youfen Xu , Ganghua Han , Hongxun Zhang , Zhisheng Yu , Ruyin Liu
J. Microbiol. 2022;60(6):594-601.   Published online April 18, 2022
DOI: https://doi.org/10.1007/s12275-022-1651-9
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  • 9 Web of Science
  • 9 Crossref
AbstractAbstract PDF
Community-based microbial source tracking (MST) can be used to determine fecal contamination from multiple sources in the aquatic environment. However, there is little scientific information on its application potential in water environmental management. Here, we compared SourceTracker and Fast Expectation-maximization Microbial Source Tracking (FEAST) performances on environmental water bodies exposed to low fecal pollution and evaluated treatment effects of fecal pollution in the watershed utilizing community-based MST. Our results showed that FEAST overall outperformed SourceTracker in sensitivity and stability, and was able to discern multi-source fecal contamination (mainly chicken feces) in ambient water bodies exposed to low fecal inputs. Consistent with our previous PCR/qPCR-based MST assays, FEAST analysis indicates that fecal pollution has been significantly mitigated through comprehensive environmental treatment by the local government. This study suggests that FEAST can be a powerful tool for accurately evaluating the contribution of multi-source fecal contamination in environmental water, facilitating environmental management.

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  • Novel Microbial Engraftment Trajectories Following Microbiota Transplant Therapy in Ulcerative Colitis
    Daphne Moutsoglou, Aneesh Syal, Sharon Lopez, Elizabeth C Nelson, Lulu Chen, Amanda J Kabage, Monika Fischer, Alexander Khoruts, Byron P Vaughn, Christopher Staley
    Journal of Crohn's and Colitis.2025;[Epub]     CrossRef
  • Integrating microbial community dynamics and emerging contaminants (ECs) for precisely quantifying the sources in groundwater affected by livestock farming
    Kai Liu, Jinrong Qiu, Chih-Huang Weng, Zhongen Tang, Renchuan Fu, Xiaojun Lin, Xiujuan Wang, Na Liu, Jingwen Zeng
    Journal of Hazardous Materials.2025; 494: 138691.     CrossRef
  • SourceApp: A Novel Metagenomic Source Tracking Tool that can Distinguish between Fecal Microbiomes Using Genome-To-Source Associations Benchmarked Against Mixed Input Spike-In Mesocosms
    Blake G. Lindner, Katherine E. Graham, Jacob R. Phaneuf, Janet K. Hatt, Konstantinos T. Konstantinidis
    Environmental Science & Technology.2025; 59(19): 9507.     CrossRef
  • A Practical Framework for Environmental Antibiotic Resistance Monitoring in Freshwater Ecosystems
    Irene Beltrán de Heredia, Itziar Alkorta, Carlos Garbisu, Estilita Ruiz-Romera
    Antibiotics.2025; 14(8): 840.     CrossRef
  • Maternal–to–neonatal microbial transmission and impact of prenatal probiotics on neonatal gut development
    Lulu Meng, Ge Fan, Haishan Xie, Kian Deng Tye, Lianyi Xia, Huijuan Luo, Xiaomei Tang, Ting Huang, Jiaxin Lin, Guangyu Ma, Xiaomin Xiao, Zhe Li
    Journal of Translational Medicine.2025;[Epub]     CrossRef
  • Faecal source apportionment using molecular methods: A proof of concept using the FEAST algorithm
    Laura T. Kelly, Jack Sissons, Lucy Thompson, John K. Pearman
    Water Research.2024; 266: 122365.     CrossRef
  • Computational methods and challenges in analyzing intratumoral microbiome data
    Qi Wang, Zhaoqian Liu, Anjun Ma, Zihai Li, Bingqiang Liu, Qin Ma
    Trends in Microbiology.2023; 31(7): 707.     CrossRef
  • Response and recovery mechanisms of river microorganisms to gradient concentrations of estrogen
    Dan Qin, Yan Li, Nengwang Chen, Anyi Hu, Chang-Ping Yu
    Frontiers in Microbiology.2023;[Epub]     CrossRef
  • Improving the Identification of Fecal Contamination in Recreational Water through the Standardization and Normalization of Microbial Source Tracking
    Megan N. Jamison, John J. Hart, David C. Szlag
    ACS ES&T Water.2022; 2(12): 2305.     CrossRef
Regulator of ribonuclease activity modulates the pathogenicity of Vibrio vulnificus
Jaejin Lee , Eunkyoung Shin , Jaeyeong Park , Minho Lee , Kangseok Lee
J. Microbiol. 2021;59(12):1133-1141.   Published online November 9, 2021
DOI: https://doi.org/10.1007/s12275-021-1518-5
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AbstractAbstract PDF
RraA, a protein regulator of RNase E activity, plays a unique role in modulating the mRNA abundance in Escherichia coli. The marine pathogenic bacterium Vibrio vulnificus also possesses homologs of RNase E (VvRNase E) and RraA (VvRraA1 and VvRraA2). However, their physiological roles have not yet been investigated. In this study, we demonstrated that VvRraA1 expression levels affect the pathogenicity of V. vulnificus. Compared to the wild-type strain, the VvrraA1-deleted strain (ΔVvrraA1) showed decreased motility, invasiveness, biofilm formation ability as well as virulence in mice; these phenotypic changes of ΔVvrraA1 were restored by the exogenous expression of VvrraA1. Transcriptomic analysis indicated that VvRraA1 expression levels affect the abundance of a large number of mRNA species. Among them, the halflives of mRNA species encoding virulence factors (e.g., smcR and htpG) that have been previously shown to affect VvrraA1 expression-dependent phenotypes were positively correlated with VvrraA1 expression levels. These findings suggest that VvRraA1 modulates the pathogenicity of V. vulnificus by regulating the abundance of a subset of mRNA species.

Citations

Citations to this article as recorded by  
  • Identification of the global regulatory roles of RraA via the integrative transcriptome and proteome in Vibrio alginolyticus
    Huizhen Chen, Qian Gao, Bing Liu, Ying Zhang, Jianxiang Fang, Songbiao Wang, Youqi Chen, Chang Chen, Nicolas E. Buchler
    mSphere.2024;[Epub]     CrossRef
  • Comparative Transcriptomic Analysis of Flagellar-Associated Genes in Salmonella Typhimurium and Its rnc Mutant
    Seungmok Han, Ji-Won Byun, Minho Lee
    Journal of Microbiology.2024; 62(1): 33.     CrossRef
  • Eco-Evolutionary Drivers of Vibrio parahaemolyticus Sequence Type 3 Expansion: Retrospective Machine Learning Approach
    Amy Marie Campbell, Chris Hauton, Ronny van Aerle, Jaime Martinez-Urtaza
    JMIR Bioinformatics and Biotechnology.2024; 5: e62747.     CrossRef
  • Relaxed Cleavage Specificity of Hyperactive Variants of Escherichia coli RNase E on RNA I
    Dayeong Bae, Hana Hyeon, Eunkyoung Shin, Ji-Hyun Yeom, Kangseok Lee
    Journal of Microbiology.2023; 61(2): 211.     CrossRef
  • Regulator of RNase E activity modulates the pathogenicity of Salmonella Typhimurium
    Jaejin Lee, Eunkyoung Shin, Ji-Hyun Yeom, Jaeyoung Park, Sunwoo Kim, Minho Lee, Kangseok Lee
    Microbial Pathogenesis.2022; 165: 105460.     CrossRef
iTRAQ-facilitated proteomic analysis of Bacillus cereus via degradation of malachite green
Bobo Wang , Jing Lu , Junfang Zheng , Zhisheng Yu
J. Microbiol. 2021;59(2):142-150.   Published online February 1, 2021
DOI: https://doi.org/10.1007/s12275-021-0441-0
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AbstractAbstract PDF
The wide use of malachite green (MG) as a dye has caused substantial concern owing to its toxicity. Bacillus cereus can against the toxic effect of MG and efficiently decolourise it. However, detailed information regarding its underlying adaptation and degradation mechanisms based on proteomic data is scarce. In this study, the isobaric tags for relative and absolute quantitation (iTRAQ)-facilitated quantitative method was applied to analyse the molecular mechanisms by which B. cereus degrades MG. Based on this analysis, 209 upregulated proteins and 198 downregulated proteins were identified with a false discovery rate of 1% or less during MG biodegradation. Gene ontology and KEGG analysis determined that the differentially expressed proteins were enriched in metabolic processes, catalytic activity, antioxidant activity, and responses to stimuli. Furthermore, real-time qPCR was utilised to further confirm the regulated proteins involved in benzoate degradation. The proteins BCE_4076 (Acetyl-CoA acetyltransferase), BCE_5143 (Acetyl-CoA acetyltransferase), BCE_5144 (3-hydroxyacyl-CoA dehydrogenase), BCE_4651 (Enoyl-CoA hydratase), and BCE_5474 (3-hydroxyacyl-CoA dehydrogenase) involved in the benzoate degradation pathway may play an important role in the biodegradation of MG by B. cereus. The results of this study not only provide a comprehensive view of proteomic changes in B. cereus upon MG loading but also shed light on the mechanism underlying MG biodegradation by B. cereus.

Citations

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  • Engineering globins for efficient biodegradation of malachite green: two case studies of myoglobin and neuroglobin
    Jiao Liu, Jia-Kun Xu, Hong Yuan, Xiao-Juan Wang, Shu-Qin Gao, Ge-Bo Wen, Xiang-Shi Tan, Ying-Wu Lin
    RSC Advances.2022; 12(29): 18654.     CrossRef
The putative polysaccharide synthase AfCps1 regulates Aspergillus fumigatus morphogenesis and conidia immune response in mouse bone marrow-derived macrophages
Sha Wang , Anjie Yuan , Liping Zeng , Sikai Hou , Meng Wang , Lei Li , Zhendong Cai , Guowei Zhong
J. Microbiol. 2021;59(1):64-75.   Published online November 17, 2020
DOI: https://doi.org/10.1007/s12275-021-0347-x
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AbstractAbstract PDF
Aspergillus fumigatus is a well-known opportunistic pathogen that causes invasive aspergillosis (IA) infections with high mortality in immunosuppressed individuals. Morphogenesis, including hyphal growth, conidiation, and cell wall biosynthesis is crucial in A. fumigatus pathogenesis. Based on a previous random insertional mutagenesis library, we identified the putative polysaccharide synthase gene Afcps1 and its paralog Afcps2. Homologs of the cps gene are commonly found in the genomes of most fungal and some bacterial pathogens. Afcps1/cpsA is important in sporulation, cell wall composition, and virulence. However, the precise regulation patterns of cell wall integrity by Afcps1/cpsA and further effects on the immune response are poorly understood. Specifically, our in-depth study revealed that Afcps1 affects cell-wall stability, showing an increased resistance of ΔAfcps1 to the chitinmicrofibril destabilizing compound calcofluor white (CFW) and susceptibility of ΔAfcps1 to the β-(1,3)-glucan synthase inhibitor echinocandin caspofungin (CS). Additionally, deletion of Afcps2 had a normal sporulation phenotype but caused hypersensitivity to Na+ stress, CFW, and Congo red (CR). Specifically, quantitative analysis of cell wall composition using high-performance anion exchange chromatography- pulsed amperometric detector (HPAEC-PAD) analysis revealed that depletion of Afcps1 reduced cell wall glucan and chitin contents, which was consistent with the downregulation of expression of the corresponding biosynthesis genes. Moreover, an elevated immune response stimulated by conidia of the ΔAfcps1 mutant in marrow-derived macrophages (BMMs) during phagocytosis was observed. Thus, our study provided new insights into the function of polysaccharide synthase Cps1, which is necessary for the maintenance of cell wall stability and the adaptation of conidia to the immune response of macrophages in A. fumigatus.

Citations

Citations to this article as recorded by  
  • Computational Approaches for Discovering Virulence Factors in Coccidioides
    Arianna D. Daniel, Vikram Senthil, Katrina K. Hoyer
    Journal of Fungi.2025; 11(10): 754.     CrossRef
  • Study on the metabolic changes and regulatory mechanism of Aspergillus flavus conidia germination
    Sifan Jia, Chong Li, Yu An, Desheng Qi, Erik F. Y. Hom
    Microbiology Spectrum.2024;[Epub]     CrossRef
  • Chitin Biosynthesis in Aspergillus Species
    Veronica S. Brauer, André M. Pessoni, Mateus S. Freitas, Marinaldo P. Cavalcanti-Neto, Laure N. A. Ries, Fausto Almeida
    Journal of Fungi.2023; 9(1): 89.     CrossRef
  • Evidencing New Roles for the Glycosyl-Transferase Cps1 in the Phytopathogenic Fungus Botrytis cinerea
    Matthieu Blandenet, Isabelle R. Gonçalves, Christine Rascle, Jean-William Dupuy, François-Xavier Gillet, Nathalie Poussereau, Mathias Choquer, Christophe Bruel
    Journal of Fungi.2022; 8(9): 899.     CrossRef
Vibrio parahaemolyticus cqsA controls production of quorum sensing signal molecule 3-hydroxyundecan-4-one and regulatessensing signal molecule 3-hydroxyundecan-4-one and regulates colony morphology
Kui Wu , Yangyun Zheng , Qingping Wu , Haiying Chen , Songzhe Fu , Biao Kan , Yongyan Long , Xiansheng Ni , Junling Tu
J. Microbiol. 2019;57(12):1105-1114.   Published online November 4, 2019
DOI: https://doi.org/10.1007/s12275-019-9379-x
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  • 11 Crossref
AbstractAbstract PDF
In order to adapt to different environments, Vibrio parahaemolyticus employed a complicated quorum sensing system to orchestrate gene expression and diverse colony morphology patterns. In this study, the function of the putative quorum sensing signal synthase gene cqsA (VPA0711 in V. parahaemolyticus strain RIMD2210633 genome) was investigated. The cloning and expression of V. parahaemolyticus cqsA in Escherichia coli system induced the production of a new quorum sensing signal that was found in its culture supernatant. The signal was purified by high performance liquid chromatography
methods
and determined to be 3-hydroxyundecan- 4-one by indirect and direct mass spectra assays. The deletion of cqsA in RIMD2210633 changed V. parahaemolyticus colony morphology from the classical ‘fried-egg’ shape (thick and opaque in the center, while thin and translucent in the edge) of the wild-type colony to a ‘pancake’ shape (no significant difference between the centre and the edge) of the cqsAdeleted colony. This morphological change could be restored by complementary experiment with cqsA gene or the signal extract. In addition, the expression of opaR, a well-known quorum sensing regulatory gene, could be up-regulated by cqsA deletion. Our results suggested that V. parahaemolyticus used cqsA to produce 3-hydroxyundecan-4-one signal and thereby regulated colony morphology and other quorum sensing-associated behaviors.

Citations

Citations to this article as recorded by  
  • Antimicrobial resistance, virulence factors and phylogenetic profiles of Vibrio parahaemolyticus in the eastern coast of Shenzhen
    Xian Qiang Lian, Guo Dong Liu, Miao Fen Huang, Qiu Hua Fan, Zi Dan Lin
    Frontiers in Microbiology.2024;[Epub]     CrossRef
  • Quorum sensing signal synthases enhance Vibrio parahaemolyticus swarming motility
    Fuwen Liu, Fei Wang, Yixuan Yuan, Xiaoran Li, Xiaojun Zhong, Menghua Yang
    Molecular Microbiology.2023; 120(2): 241.     CrossRef
  • Regulation of Virulence Factors Expression During the Intestinal Colonization of Vibrio parahaemolyticus
    Jingyu Wang, Yuming Zhan, Han Sun, Xiaodan Fu, Qing Kong, Changliang Zhu, Haijin Mou
    Foodborne Pathogens and Disease.2022; 19(3): 169.     CrossRef
  • Supplementation of ex situ produced bioflocs improves immune response against AHPND in Pacific whiteleg shrimp (Litopenaeus vannamei) postlarvae
    Magdalena Lenny Situmorang, Umaporn Uawisetwathana, Sopacha Arayamethakorn, Nitsara Karoonuthaisiri, Wanilada Rungrassamee, Haniswita Haniswita, Peter Bossier, Gede Suantika
    Applied Microbiology and Biotechnology.2022; 106(9-10): 3751.     CrossRef
  • A novel finding of intra-genus inhibition of quorum sensing in Vibrio bacteria
    Huong Thanh Hoang, Thuy Thu Thi Nguyen, Ha Minh Do, Thao Kim Nu Nguyen, Hai The Pham
    Scientific Reports.2022;[Epub]     CrossRef
  • CqsA-introduced quorum sensing inhibits type VI secretion system 2 through an OpaR-dependent pathway in Vibrio parahaemolyticus
    Kui Wu, Yongyan Long, Qian Liu, Wei Wang, Guoyin Fan, Hui Long, Yangyun Zheng, Xiansheng Ni, Shengen Chen, Haiying Chen, Shufen Shuai
    Microbial Pathogenesis.2022; 162: 105334.     CrossRef
  • CqsA inhibits the virulence of Vibrio harveyi to the pearl gentian grouper (♀Epinephelus fuscoguttatus × ♂Epinephelus lanceolatus)
    Yaqiu Zhang, Yiqin Deng, Juan Feng, Zhixun Guo, Can Mao, Haoxiang Chen, Ziyang Lin, Jianmei Hu, Youlu Su
    Aquaculture.2021; 535: 736346.     CrossRef
  • Identification of LuxR Family Regulators That Integrate Into Quorum Sensing Circuit in Vibrio parahaemolyticus
    Xiaojun Zhong, Ranran Lu, Fuwen Liu, Jinjie Ye, Junyang Zhao, Fei Wang, Menghua Yang
    Frontiers in Microbiology.2021;[Epub]     CrossRef
  • Adaptations of Vibrio parahaemolyticus to Stress During Environmental Survival, Host Colonization, and Infection
    Gururaja Perumal Pazhani, Goutam Chowdhury, Thandavarayan Ramamurthy
    Frontiers in Microbiology.2021;[Epub]     CrossRef
  • Vibrio alginolyticus influences quorum sensing-controlled phenotypes of acute hepatopancreatic necrosis disease-causing Vibrio parahaemolyticus
    Panida Paopradit, Natta Tansila, Komwit Surachat, Pimonsri Mittraparp-arthorn
    PeerJ.2021; 9: e11567.     CrossRef
  • Dynamics and Microevolution of Vibrio parahaemolyticus Populations in Shellfish Farms
    Songzhe Fu, Qingyao Wang, Yixiang Zhang, Qian Yang, Jingwei Hao, Ying Liu, Bo Pang, Michael S. Rappe
    mSystems.2021;[Epub]     CrossRef
Two novel synthetic peptides inhibit quorum sensing-dependent biofilm formation and some virulence factors in Pseudomonas aeruginosa PAO1
Mostafa N. Taha , Amal E. Saafan , A. Ahmedy , Eman El Gebaly , Ahmed S. Khairalla
J. Microbiol. 2019;57(7):618-625.   Published online June 27, 2019
DOI: https://doi.org/10.1007/s12275-019-8548-2
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AbstractAbstract PDF
Quorum sensing (QS) regulates virulence factor expression in Pseudomonas aeruginosa. Inhibiting the QS-controlled virulence factors without inhibiting the growth of P. aeruginosa is a promising approach for overcoming the widespread resistance of P. aeruginosa. This study was proposed to investigate the effects of two novel synthetic peptides on the biofilm development and virulence factor production of P. aeruginosa. The tested strain was P. aeruginosa PAO1. The results indicated that both of the synthetic peptides (LIVRHK and LIVRRK) inhibited (P < 0.05) the formation of biofilms and the production of virulence factors, including pyocyanin, protease, and rhamnolipids, without inhibiting the growth of PAO1. Additionally, we detected transcriptional changes related to QS and found a significant reduction in the levels of gene expression of lasI, lasR, rhlI, and rhlR. This study demonstrates that LIVRRK and LIVRHK are novel synthetic peptides that can act as potent inhibitors of QS-regulated virulence factors in P. aeruginosa. Moreover, these synthetic peptides have potential applications in the treatment of biofilmrelated diseases. Both peptides may be able to control chronic infections and biofilm-associated problems of P. aeruginosa.

Citations

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  • Peptide-based approaches to quorum-sensing disruption: emerging trends and applications in antimicrobial therapy
    Mo Ahamad Khan, Lechen Zhu, Hu Zhu
    Bioorganic & Medicinal Chemistry.2026; 133: 118496.     CrossRef
  • Novel quorum-sensing inhibitor peptide SF derived from Penaeus vannamei myosin inhibits biofilm formation and virulence factors in Vibrio parahaemolyticus
    Wenqi Yang, Shuilin Liu, Xueqing Liu, Shuo Yuan, Le An, Anqi Ren, Fengling Bai, Xinran Lv, Jianrong Li, Xuepeng Li, Yuqiong Meng, Rui Ma
    LWT.2025; 218: 117542.     CrossRef
  • Harnessing Chemical Diversity to Disarm Bacterial Communication for 25 Years: Natural Products, Repurposed Drugs, Peptides, and Synthetic Quorum Sensing Inhibitors
    Tung Truong‐Thanh, Anh Nguyen Phuong, Linh Nguyen Khanh, Duc Nguyen Minh
    ChemMedChem.2025;[Epub]     CrossRef
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    Abdelaziz Touati, Nasir Adam Ibrahim, Lilia Tighilt, Takfarinas Idres
    Microorganisms.2025; 13(8): 1838.     CrossRef
  • Rational Design, Computational Analysis and Antibacterial Activities of Synthesized Peptide-Based Molecules Targeting Quorum Sensing-Dependent Biofilm Formation in Pseudomonas aeruginosa
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    Pharmaceuticals.2025; 18(10): 1572.     CrossRef
  • Biofilm prevention and quorum sensing interference via surface-bound peptoid
    Francesca Gamna, Andrea Cochis, Gustavo Penteado Battesini Carretero, Jovana Curcic, Biljana Mojsoska, Milka Malesevic, Ziba Najmi, Lia Rimondini, Silvia Spriano
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    Jiarui Du, Jiahao Li, Juqi Wen, Jun Liu, Haichuan Xiao, Antian Zhang, Dongdong Yang, Pinghua Sun, Haibo Zhou, Jun Xu
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    Bioorganic Chemistry.2023; 141: 106922.     CrossRef
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    Yangrui Wang, Mengtong Sun, Xiaoling Cui, Yongyue Gao, Xinran Lv, Jianrong Li, Fengling Bai, Xuepeng Li, Defu Zhang, Kai Zhou
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    Fatemeh Aflakian, Mehrnaz Rad, Gholamreza Hashemitabar, Milad Lagzian, Mohammad Ramezani
    Biofouling.2022; 38(2): 131.     CrossRef
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    Israel Castillo-Juárez, Blanca Esther Blancas-Luciano, Rodolfo García-Contreras, Ana María Fernández-Presas
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    Journal of Agricultural and Food Chemistry.2022; 70(25): 7716.     CrossRef
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    Biophysical Reviews.2021; 13(1): 35.     CrossRef
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    MicrobiologyOpen.2020;[Epub]     CrossRef
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    Jiarui Li, Pablo Fernández-Millán, Ester Boix
    Current Topics in Medicinal Chemistry.2020; 20(14): 1238.     CrossRef
  • Olive Leaf Extract Modulates Quorum Sensing Genes and Biofilm Formation in Multi-Drug Resistant Pseudomonas aeruginosa
    Nazly R. El-sayed, Reham Samir, Lina Jamil M. Abdel-Hafez, Mohammed A. Ramadan
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A common evolutionary pathway for maintaining quorum sensing in Pseudomonas aeruginosa
Bai-min Lai , Hui-cong Yan , Mei-zhen Wang , Na Li , Dong-sheng Shen
J. Microbiol. 2018;56(2):83-89.   Published online February 2, 2018
DOI: https://doi.org/10.1007/s12275-018-7286-1
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AbstractAbstract PDF
In the bacterium Pseudomonas aeruginosa, the synthesis and secretion of extracellular protease is a typical cooperative behavior regulated by quorum sensing. However, this type of cooperative behavior is easily exploited by other individuals who do not synthesize public goods, which is known as the “tragedy of the commons”. Here P. aeruginosa was inoculated into casein media with different nitrogen salts added. In casein broth, protease (a type of public good) is necessary for bacterial growth. After 30 days of sequential transfer, some groups propagated stably and avoided “tragedy of the commons”. The evolved cooperators who continued to synthesize protease were isolated from these stable groups. By comparing the characteristics of quorum sensing in these cooperators, an identical evolutionary pattern was found. A variety of cooperative behaviors regulated by quorum sensing, such as the synthesis and secretion of protease and signals, were significantly reduced during the process of evolution. Such reductions improved the efficiency of cooperation, helping to prevent cheating. In addition, the production of pyocyanin, which is regulated by the RhlIR system, increased during the process of evolution, possibly due to its role in stabilizing the cooperation. This study contributes towards our understanding of the evolution of quorum sensing of P. aeruginosa.

Citations

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  • Role of horizontal gene transfer and cooperation in rhizosphere microbiome assembly
    Simone Raposo Cotta, Armando Cavalcante Franco Dias, Rodrigo Mendes, Fernando Dini Andreote
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Mutation of the cyclic di-GMP phosphodiesterase gene in Burkholderia lata SK875 attenuates virulence and enhances biofilm formation
Hae-In Jung , Yun-Jung Kim , Yun-Jung Lee , Hee-Soo Lee , Jung-Kee Lee , Soo-Ki Kim
J. Microbiol. 2017;55(10):800-808.   Published online September 28, 2017
DOI: https://doi.org/10.1007/s12275-017-7374-7
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AbstractAbstract PDF
Burkholderia sp. is a gram-negative bacterium that commonly exists in the environment, and can cause diseases in plants, animals, and humans. Here, a transposon mutant library of a Burkholderia lata isolate from a pig with swine respiratory disease in Korea was screened for strains showing attenuated virulence in Caenorhabditis elegans. One such mutant was obtained, and the Tn5 insertion junction was mapped to rpfR, a gene encoding a cyclic di-GMP phosphodiesterase that functions as a receptor. Mutation of rpfR caused a reduction in growth on CPG agar and swimming motility as well as a rough colony morphology on Congo red agar. TLC analysis showed reduced AHL secretion, which was in agreement with the results from plate-based and bioluminescence assays. The mutant strain produced significantly more biofilm detected by crystal violet staining than the parent strain. SEM of the mutant strain clearly showed that the overproduced biofilm contained a filamentous structure. These results suggest that the cyclic di-GMP phosphodiesterase RpfR plays an important role in quorum sensing modulation of the bacterial virulence and biofilm formation.

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  • Functional analysis of quorum sensing-mediated pathogenicity in Burkholderia contaminans SK875 using transposon mutagenesis
    Kai-Min Niu, Yun Jung Lee, Hae-In Jung, Damini Kothari, Digar Singh, Soo-Ki Kim
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  • Comprehensive genome analysis of Burkholderia contaminans SK875, a quorum-sensing strain isolated from the swine
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  • Comparative genomics and transcriptomic response to root exudates of six rice root-associated Burkholderia sensu lato species
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D-Galactose as an autoinducer 2 inhibitor to control the biofilm formation of periodontopathogens
Eun-Ju Ryu , Jaehyun Sim , Jun Sim , Julian Lee , Bong-Kyu Choi
J. Microbiol. 2016;54(9):632-637.   Published online August 31, 2016
DOI: https://doi.org/10.1007/s12275-016-6345-8
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AbstractAbstract PDF
Autoinducer 2 (AI-2) is a quorum sensing molecule to which bacteria respond to regulate various phenotypes, including virulence and biofilm formation. AI-2 plays an important role in the formation of a subgingival biofilm composed mostly of Gram-negative anaerobes, by which periodontitis is initiated. The aim of this study was to evaluate D-galactose as an inhibitor of AI-2 activity and thus of the biofilm formation of periodontopathogens. In a search for an AI-2 receptor of Fusobacterium nucleatum, D-galactose binding protein (Gbp, Gene ID FN1165) showed high sequence similarity with the ribose binding protein (RbsB), a known AI-2 receptor of Aggregatibacter actinomycetemcomitans. D-Galactose was evaluated for its inhibitory effect on the AI-2 activity of Vibrio harveyi BB152 and F. nucleatum, the major coaggregation bridge organism, which connects early colonizing commensals and late pathogenic colonizers in dental biofilms. The inhibitory effect of D-galactose on the biofilm formation of periodontopathogens was assessed by crystal violet staining and confocal laser scanning microscopy in the absence or presence of AI-2 and secreted molecules of F. nucleatum. D-Galactose significantly inhibited the AI-2 activity of V. harveyi and F. nucleatum. In addition, D-galactose markedly inhibited the biofilm formation of F. nucleatum, Porphyromonas gingivalis, and Tannerella forsythia induced by the AI-2 of F. nucleatum without affecting bacterial growth. Our
results
demonstrate that the Gbp may function as an AI-2 receptor and that galactose may be used for prevention of the biofilm formation of periodontopathogens by targeting AI-2 activity.

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    Jae‐Young Byeon, Jun Sim, Eun‐Ju Ryu, Jaehyun Sim, Hwan Lee, Kwang‐Hwi Cho, Bong‐Kyu Choi, Julian Lee
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    Jeffrey L. Ebersole, Dolph Dawson, Pinar Emecen‐Huja, Radhakrishnan Nagarajan, Katherine Howard, Martha E. Grady, Katherine Thompson, Rebecca Peyyala, Ahmad Al‐Attar, Kathryn Lethbridge, Sreenatha Kirakodu, Octavio A. Gonzalez
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Research Support, Non-U.S. Gov'ts
D101 is critical for the function of AttJ, a repressor of quorum quenching system in Agrobacterium tumefaciens
Chao Wang , Chunlan Yan , Yong-Gui Gao , Lian-Hui Zhang
J. Microbiol. 2015;53(9):623-632.   Published online August 1, 2015
DOI: https://doi.org/10.1007/s12275-015-5100-x
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AbstractAbstract PDF
The quorum quenching system of Agrobacterium tumefaciens is specifically activated upon entering the stationary phase. Evidence has shown that this system includes two key components: the IclR-type transcriptional factor AttJ (also named as BlcR) and the AHL-lactonase AttM (also named as BlcC). At exponential phase, AttJ binds to the promoter region of attM and thus suppresses the expression of attM. At stationary phase, however, the small molecule SSA directly binds to AttJ and relieves its inhibition of AttJ and thereby triggers the expression of attM. While the regulation of AttM has been extensively investigated, little is known about the regulation of AttJ. In this study, we demonstrated the D101 amino acid of AttJ is essential for the AttJ function. In vitro, the variant protein of AttJD101H appeared to be readily aggregated. In vivo, the D101H mutation in AttJ entirely abolished the inhibitory activity of AttJ and overexpressed attM in A. tumefaciens A6. In addition, D101H mutation led to an overexpression of attJ, indicating an auto-regulatory mechanism for the attJ regulation. Put together, these findings demonstrate that D101 is an important amino acid for the transcription activity of AttJ and the transcription of attJ is regulated by a negative feedback loop. These results expand previous biochemical characterization of AttJ and provide new mechanistic insights into the regulation of quorum quenching in A. tumefaciens.
Note] Inhibition of quorum sensing in Pseudomonas aeruginosa by two herbal essential oils from Apiaceae family
Ehsan Sepahi , Saeed Tarighi , Farajollah Shahriari Ahmadi , Abdolreza Bagheri
J. Microbiol. 2015;53(2):176-180.   Published online January 5, 2015
DOI: https://doi.org/10.1007/s12275-015-4203-8
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AbstractAbstract
Ferula (Ferula asafoetida L.) and Dorema (Dorema aucheri Bioss.) both from Apiaceae family were tested for their antiquorum sensing (QS) activity against Pseudomonas aeruginosa. Both essential oils exhibited anti-QS activity at 25 μg/ml of concenteration. At this concenteration Ferula fully abolished and Dorema reduced the violacein production by C. violaceum. Pyocyanin, pyoverdine, elastase and biofilm production were decreased in Ferula oil treatments. Dorema oil reduced pyoverdine and elastase production, while pyocyanin and biofilm production were not affacted. Expresion analysis of QS-dependent genes confirmed our phenotypic data. Our data introduced native Dorema and Ferula plants as novel QS and virulence inhibitors.

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Inhibitory Effects of 4-Hydroxy-2,5-Dimethyl-3(2H)-Furanone (HDMF) on Acyl-Homoserine Lactone-Mediated Virulence Factor Production and Biofilm Formation in Pseudomonas aeruginosa PAO1
Sung-Chan Choi , Can Zhang , Sooyoung Moon , Young-Sook Oh
J. Microbiol. 2014;52(9):734-742.   Published online August 2, 2014
DOI: https://doi.org/10.1007/s12275-014-4060-x
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AbstractAbstract PDF
4-Hydroxy-2,5-dimethyl-3(2H)-furanone (HDMF), a nonhalogenated furanone found in a variety of fruits, has been shown to have antimicrobial activity. However, few studies have focused on its inhibitory effect on bacterial quorum sensing (QS) at levels below the non-inhibitory concentration. In this study, 0.1 μM HDMF decreased the production of QS signal molecules and inhibited QS-controlled biofilm formation by Pseudomonas aeruginosa PAO1 without causing growth inhibition. In the presence of 0.1 and 1.0 μM HDMF, biofilm production by PAO1 was reduced by 27.8 and 42.6%, respectively, compared to that by untreated control cells. HDMF (1.0 μM) also significantly affected virulence factor expression (regulated by the las, rhl, and pqs system), resulting in a significant reduction in the production of LasA protease (53.8%), rhamnolipid (40.9%), and pyocyanin (51.4%). This HDMF-dependent inhibition of virulence factor expression was overcome by increasing the levels of two QS signal molecules of P. aeruginosa, N-(3-oxo-dodecanoyl)-L-homoserine lactone and N-butyryl-L-homoserine lactone, suggesting reversible competitive inhibition between HDMF and these molecules. The results of this study indicate that HDMF has great potential as an inhibitor of QS, and that it may be of value as a therapeutic agent and in biofilm control, without increasing selective pressure for resistance development.

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A Functional and Phylogenetic Comparison of Quorum Sensing Related Genes in Brucella melitensis 16M
Aniel Jessica Leticia Brambila-Tapia , Ernesto Pérez-Rueda
J. Microbiol. 2014;52(8):709-715.   Published online July 4, 2014
DOI: https://doi.org/10.1007/s12275-014-3570-x
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AbstractAbstract PDF
A quorum-sensing (QS) system is involved in Brucella melitensis survival inside the host cell. Two transcriptional regulators identified in B. melitensis, BlxR and VjbR, regulate the expression of virB, an operon required for bacterial intracellular persistence. In this work, 628 genes affected by VjbR and 124 by BlxR were analyzed to gain insights into their functional and taxonomical distributions among the Bacteria and Archaea cellular domains. In this regard, the Cluster of Orthologous Groups (COG) genes and orthologous genes in 789 nonredundant bacterial and archaeal genomes were obtained and compared against a group of randomly selected genes. From these analyses, we found 71 coaffected genes between VjbR and BlxR. In the COG comparison, VjbR activated genes associated with intracellular trafficking, secretion and vesicular transport and defense mechanisms, while BlxR affected genes related to energy production and conversion (with an equal effect) and translation, ribosomal structure and biogenesis, posttranslational modifications and carbohydrate and amino acid metabolism (with a negative effect). When the taxonomical distribution of orthologous genes was evaluated, the VjbR- and BlxRrelated genes presented more orthologous genes in Crenarchaeota (Archaea), Firmicutes, and Tenericutes and fewer genes in Proteobacteria than expected by chance. These findings suggest that QS system exert a fine-tuning modulation of gene expression, by which VjbR activates genes related to infection persistence and defense, while BlxR represses general bacterial metabolism for intracellular adaptations. Finally, these affected genes present a degree of presence among Bacteria and Archaea genomes that is different from that expected by chance.

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Review
REVIEW] Mechanisms of Synergy in Polymicrobial Infections
Justine L. Murray , Jodi L. Connell , Apollo Stacy , Keith H. Turner , Marvin Whiteley
J. Microbiol. 2014;52(3):188-199.   Published online March 1, 2014
DOI: https://doi.org/10.1007/s12275-014-4067-3
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AbstractAbstract PDF
Communities of microbes can live almost anywhere and contain many different species. Interactions between members of these communities often determine the state of the habitat in which they live. When these habitats include sites on the human body, these interactions can affect health and disease. Polymicrobial synergy can occur during infection, in which the combined effect of two or more microbes on disease is worse than seen with any of the individuals alone. Powerful genomic methods are increasingly used to study microbial communities, including metagenomics to reveal the members and genetic content of a community and metatranscriptomics to describe the activities of community members. Recent efforts focused toward a mechanistic understanding of these interactions have led to a better appreciation of the precise bases of polymicrobial synergy in communities containing bacteria, eukaryotic microbes, and/or viruses. These studies have benefited from advances in the development of in vivo models of polymicrobial infection and modern techniques to profile the spatial and chemical bases of intermicrobial communication. This review describes the breadth of mechanisms microbes use to interact in ways that impact pathogenesis and techniques to study polymicrobial communities.

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Research Support, Non-U.S. Gov'ts
An Aqueous Extract of Yunnan Baiyao Inhibits the Quorum-Sensing-Related Virulence of Pseudomonas aeruginosa
Zu-Guo Zhao , Shuang-Shuang Yan , Yun-Mei Yu , Na Mi , La-Xi Zhang , Jun Liu , Xiao-Ling Li , Fang Liu , Jun-Fa Xu , Wei-Qing Yang , Guo-Ming Li
J. Microbiol. 2013;51(2):207-212.   Published online April 27, 2013
DOI: https://doi.org/10.1007/s12275-013-2595-x
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AbstractAbstract PDF
Yunnan Baiyao is a famous Chinese medicine that has long been directly applied to wounds to reduce bleeding, pain, and swelling without causing infection. However, little is known about its ability to prevent infection. The present study aimed to assess in vitro the anti-virulence activity of an aqueous extract of Yunnan Baiyao (YBX) using Pseudomonas aeruginosa as a pathogenic model. We found that a sub-MIC (2.5 mg/ml) of YBX can efficiently interfere with the quorum-sensing (QS) signaling circuit. Real-time polymerase chain reaction analysis showed that a sub-MIC of YBX downregulated the transcriptions of lasR, lasI, rhlR, and rhlI, which resulted in global attenuation of QS-regulated virulence activities, such as biofilm formation, and secretion of LasA protease, LasB elastase and pyocyanin. Further, YBX reduced the motility of P. aeruginosa related to QS, and impaired the formation of biofilms. These results suggest that YBX may possess global inhibitory activity against the virulence of P. aeruginosa and that YBX may also exhibit antimicrobial activity in vivo. The present study suggests that Yunnan Baiyao represents a potential source for isolating novel, safe, and efficacious antimicrobial agents.
A New Quorum-Sensing Inhibitor Attenuates Virulence and Decreases Antibiotic Resistance in Pseudomonas aeruginosa
Yu-Xiang Yang , Zhen-Hua Xu , Yu-Qian Zhang , Jing Tian , Li-Xing Weng , Lian-Hui Wang
J. Microbiol. 2012;50(6):987-993.   Published online December 30, 2012
DOI: https://doi.org/10.1007/s12275-012-2149-7
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AbstractAbstract PDF
Quorum sensing (QS) has been a novel target for the treatment of infectious diseases. Here structural analogs of Pseudomonas aeruginosa autoinducer N-acyl homoserine lactone (AHL) were investigated for QS inhibitor (QSI) activity and a novel QSI was discovered, N-decanoyl-L-homoserine benzyl ester (C2). Virulence assays showed that C2 downregulated total protease and elastase activities, as well as the production of rhamnolipid, that are controlled by QS in P. aeruginosa wild-type strain PAO1 without affecting growth. C2 was also shown to inhibit swarming motility of PAO1. Using a microdilution checkerboard method, we identified synergistic interactions between C2 and several antibiotics, tobramycin, gentamycin, cefepime, and meropenem. Data from real-time RT-PCR suggested that C2 inhibited the expression of lasR (29.67%), lasI (21.57%), rhlR (28.20%), and rhlI (29.03%).

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Functional Definition of LuxS, an Autoinducer-2 (AI-2) Synthase and Its Role in Full Virulence of Streptococcus suis Serotype 2
Min Cao , Youjun Feng , Changjun Wang , Feng Zheng , Ming Li , Hui Liao , Yinghua Mao , Xiuzhen Pan , Jing Wang , Dan Hu , Fuquan Hu , Jiaqi Tang
J. Microbiol. 2011;49(6):1000-1011.   Published online December 28, 2011
DOI: https://doi.org/10.1007/s12275-011-1523-1
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AbstractAbstract PDF
Quorum sensing is a widespread chemical communication in response to fluctuation of bacterial population density, and has been implicated into bacterial biofilm formation and regulation of expression of virulence factors. The luxS gene product, S-ribosylhomocysteinase, catalizes the last committed step in biosynthetic pathway of autoinducer 2 (AI-2), a signaling molecule for inter-species quorum sensing. We found a luxS homologue in 05ZYH33, an epidemic strain of Streptococcus suis serotype 2 (SS2) in China. A luxS null mutant (ΔluxS) of 05ZYH33 strain was obtained using an approach of homologous recombination. LuxS was determined to be required for AI-2 production in 05ZYH33 strain of S. suis 2. Inactivation of luxS gene led to a wide range of phenotypic changes including thinner capsular walls, increased tolerance to H2O2, reduced adherence capacity to epithelial cells, etc. In particular, loss of LuxS impaired dramatically its full virulence of SS2 in experimental model of piglets, and functional complementation restored it nearly to the level of parent strain. Genome-wide transcriptome analyses suggested that some known virulence factors such as CPS are down-regulated in the ΔluxS mutant, which might in part explain virulence attenuation by luxS deletion. Similarly, 29 of 71 genes with different expression level were proposed to be targets candidate regulated by LuxS/AI-2-dependent quorum sensing.

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Heterogeneous Virulence Potential and High Antibiotic Resistance of Pseudomonas aeruginosa Strains Isolated from Korean Pneumonia Patients
Mi Young Yoon , Kang-Mu Lee , Seok Hoon Jeong , Jungmin Kim , Sang Sun Yoon
J. Microbiol. 2010;48(4):518-525.   Published online August 20, 2010
DOI: https://doi.org/10.1007/s12275-010-9388-2
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AbstractAbstract PDF
Pseudomonas aeruginosa is an opportunistic human pathogen of clinical importance that causes airway infections in immunocompromised patients. Here, we report the virulence-associated characteristics of strains of P. aeruginosa, isolated from the sputa of 25 Korean pneumonia patients. A high degree of genomic plasticity was observed by random amplified polymorphic DNA genotype analysis, suggesting that the infections were caused by strains with diverse genomic backgrounds. Biofilm formation of each isolate was heterogeneous in terms of their relative motilities. In addition, 48% of isolates were defective in the production of 3-oxo-C12-HSL (PAI-1), a quorum sensing signal molecule. In these strains, PAI-1-dependent elastase production was correspondingly decreased, suggesting that a large number of strains were presumed to be quorum sensing deficient. Multidrug resistance (MDR) was seen in 56% of the isolates tested, and 44% of the MDR strains were resistant to five or more antibiotics. Taken together, our results provide additional insights into the virulence traits of P. aeruginosa clinical isolates, which will aid in treating P. aeruginosa infections in pneumonia patients.

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Ligand-Receptor Recognition for Activation of Quorum Sensing in Staphylococcus aureus
Li-Chun Chen , Li-Tse Tsou , Feng-Jui Chen
J. Microbiol. 2009;47(5):572-581.   Published online October 24, 2009
DOI: https://doi.org/10.1007/s12275-009-0004-2
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AbstractAbstract PDF
The accessory gene regulator (agr) locus controls many of the virulence toxins involved in Staphylococcus aureus pathogenesis, and can be divided into four specificity groups. AgrC is the only group-specific receptor to mediate both intra-group activation and inter-group inhibition. We studied the ligand-receptor recognition of the agr system in depth by using a luciferase reporter system to identify the key residues responsible for AgrC activation in two closely related agr groups, AgrC-I, and AgrC-IV. Fusion PCR and site-directed mutagenesis were used to screen for functional residues of AgrC. Our data suggest that for AgrC-IV activation, residue 101 is critical for activating the receptor. In contrast, the key residues for the activation of AgrC-I are located at residues 49~59, 107, and 116. However, three residue changes, T101A, V107S, I116S, are sufficient to convert the AIP recognizing specificity from AgrC-IV to AgrC-I.

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Acyl-Homoserine Lactone Quorum Sensing in Bacteria
E. Peter Greenberg
J. Microbiol. 2000;38(3):117-121.
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AbstractAbstract PDF
Recent advances in studies of bacterial gene expression and light microscopy show that cell-to-cell communication and community behavior are the rule rather than the exception. One type of cell-cell communication, quorum sensing in Gram-negative bacteria involves acyl-homoserine lactone signals. This type of quorum sensing represents a dedicated communication system that enables a given species to sense when it has reached a critical population density, and to respond by activating expression of specific genes. The LuxR and LuxI proteins of Vibrio fisheri are the founding members of the acyl-homoserine lactone quorum sensing signal receptor and signal generator families of proteins. Acyl-homoserine lactone signaling in Pseudomonas aeruginosa is one model for the relationship between quorum sensing, community behavior, and virulence. In the P. aeruginosa model, quorum sensing is required for normal biofilm maturation and virulence. There are multiple quorum-sensing circuits that control the expression of dozens of specific genes in P. aeruginosa.
Regulation of Class II Bacteriocin Production by Cell-Cell Signaling
Luis E. N. Quadri
J. Microbiol. 2003;41(3):175-182.
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AbstractAbstract PDF
Production of ribosomally synthesized antimicrobial peptides usually referred to as bacteriocins is an inducible trait in several gram positive bacteria, particularly in those belonging to the group of lactic acid bacteria. In many of these organisms, production of bacteriocins is inducible and induction requires secretion and extracellular accumulation of peptides that act as chemical messengers and trigger bacteriocin production. These inducer peptides are often referred to as autoinducers and are believed to permit a quorum sensing-based regulation of bacteriocin production. Notably, the peptides acting as autoinducers are dedicated peptides with or without antimicrobial activity or the bacteriocins themselves. The autoinducer-dependent induction of bacteriocin production requires histidine protein kinases and response regulator proteins of two-component signal transduction systems. The current working model for the regulation of class II bacteriocin production in lactic acid bacteria and the most relevant direct and indirect pieces of evidence supporting the model are discussed in this minireview.
Quorum Sensing and Quorum-Quenching Enzymes
Yi-Hu Dong , Lian-Hui Zhang
J. Microbiol. 2005;43(1):101-109.
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AbstractAbstract PDF
To gain maximal benefit in a competitive environment, single-celled bacteria have adopted a community genetic regulatory mechanism, known as quorum sensing (QS). Many bacteria use QS signaling systems to synchronize target gene expression and coordinate biological activities among a local population. N-acylhomoserine lactones (AHLs) are one family of the well-characterized QS signals in Gram-negative bacteria, which regulate a range of important biological functions, including virulence and biofilm formation. Several groups of AHL-degradation enzymes have recently been identified in a range of living organisms, including bacteria and eukaryotes. Expression of these enzymes in AHL-dependent pathogens and transgenic plants efficiently quenches the microbial QS signaling and blocks pathogenic infections. Discovery of these novel quorum quenching enzymes has not only provided a promising means to control bacterial infections, but also presents new challenges to investigate their roles in host organisms and their potential impacts on ecosystems.

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