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pseudomonas aeruginosa regulator PvrA binds simultaneously to multiple pseudo-palindromic sites for efficient transcription activation

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Science China(Life Sciences) 2024年第5期67卷 900-912页

作者： Yibo Zhu Bingnan Luo Xingyu Mou Yingjie Song Yonghong Zhou Yongbo Luo Bo Sun Youfu Luo Hong Tang Zhaoming Su Rui Bao Center of Infectious Diseases Division of Infectious Diseases in State Key Laboratory of BiotherapyWest China HospitalSichuan UniversityChengdu 610041China Department of Geriatrics and National Clinical Research Center for Geriatrics West China HospitalSichuan UniversityChengdu 610041China Key Laboratory of Biodiversity and Environment on the Qinghai-Tibetan Plateau Ministry of EducationSchool of Ecology and EnvironmentTibet UniversityLhasa 850000China Shanghai Advanced Research Institute Chinese Academy of SciencesShanghai 201210China College of Life Science Sichuan Normal UniversityChengdu 610101China Accurate Biotechnology(Hunan)Co. LtdChangsha 410006China

Tetracycline repressor(TetR)family regulators(TFRs)are the largest group of DNA-binding transcription factors and are widely distributed in bacteria and archaea.TFRs play vital roles in controlling the expression of various genes and regulating diverse physiological processes.Recently,a TFR protein pseudomonas virulence regulator A(PvrA),was identified from pseudomonas aeruginosa as the transcriptional activator of genes involved in fatty acid utilization and bacterial virulence.Here,we show that PvrA can simultaneously bind to multiple pseudo-palindromic sites and upregulate the expression levels of target genes.Cryo-electron microscopy(cryo-EM)analysis indicates the simultaneous DNA recognition mechanism of PvrA and suggests that the bound DNA fragments consist of a distorted B-DNA double helix.The crystal structure and functional analysis of PvrA reveal a hinge region that secures the correct domain motion for recognition of the promiscuous promoter.Additionally,our results showed that mutations disrupting the regulatory hinge region have differential effects on biofilm formation and pyocyanin biosynthesis,resulting in attenuated bacterial virulence.Collectively,these findings will improve the understanding of the relationship between the structure and function of the TetR family and provide new insights into the mechanism of regulation of P.aeruginosa virulence.

关键词： pseudomonas aeruginosa transcription regulator TetR protein-structure virulence

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RsaL is a self-regulatory switch that controls alternative biosynthesis of two AHL-type quorum sensing signals in pseudomonas aeruginosa PA1201

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mLife 2024年第1期3卷 74-86页

作者： Ya-Wen He Zi-Jing Jin Ying Cui Kai Song Bo Chen Lian Zhou State Key Laboratory of Microbial Metabolism Joint International Research Laboratory of Metabolic and Developmental SciencesSJTU-NLBP Joint R&D Centre for Biopesticides and BiofertilizersSchool of Life Sciences and BiotechnologyShanghai Jiao Tong UniversityShanghaiChina Zhiyuan Innovative Research Centre Student Innovation CentreZhiyuan CollegeShanghai Jiao Tong UniversityShanghaiChina

pseudomonas aeruginosa is a ubiquitous and metabolically versatile microorganism naturally found in soil and water.It is also an opportunistic pathogen in plants,insects,animals,and humans.In response to increasing cell density,P.aeruginosa uses two acylhomoserine lactone(AHL)quorum-sensing(QS)signals(i.e.,N-3-oxo-dodecanoyl homoserine lactone[3-oxo-C12-HSL]and Nbutanoyl-homoserine lactone[C4-HsL]),which regulate the expression of hundreds of genes.However,how the biosynthesis of these two QS signals is coordinated remains unknown.We studied the regulation of these two QS signals in the rhizosphere strain PA1201.PA1201 sequentially produced 3-oxo-C12-HSL and C4-HSL at the early and late growth stages,respectively.The highest 3-oxo-C12-HSL-dependent elastase activity was observed at the early stage,while the highest C4-HSL-dependent rhamnolipid production was observed at the late stage.The atypical regulator RsaL played a pivotal role in coordinating 3-oxo-C12-HSL and C4-HSL biosynthesis and QS-associated virulence.RsaL repressed las/transcription by binding the-10 and-35 boxes of the lasl promoter.In contrast,RsaL activated rhll transcription by binding the region encoding the 5'-untranslated region of the rhll mRNA.Further,RsaL repressed its own expression by binding a nucleotide motif located in the-35 box of the rsaL promoter.Thus,RsaL acts as a molecular switch that coordinates the sequential biosynthesis of AHL QS signals and differential virulence in PA1201.Finally,C4-HSL activation by RsaL was independent of the Las and pseudomonas quinolone signal(PQS)QS signaling systems.Therefore,we propose a new model of the QS regulatory network in PA1201,in which RsaL represents a superior player acting at the top of the hierarchy.

关键词： N-3-oxo-dodecanoyl homoserine lactone N-butanoyl-homoserine lactone pseudomonas aeruginosa quorum sensing RsaL

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pseudomonas aeruginosa infections and improper storage conditions influence the performance of 1,3‐β‐D‐glucan in diagnosis of invasive fungal infections

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iLABMED 2024年第1期2卷 53-59页

作者： Zilan Wei Jie Xu Fang Yuan Wendong Fang Jiahui Wu Youliang Wang Shuiping Chen Medical School of Chinese PLA BeijingChina Department of Laboratory Medicine 5th Medical Center of Chinese PLA General HospitalBeijingChina Department of Laboratory Medicine The PLA 307 Clinical College5th Clinical Medical College of Anhui Medical UniversityBeijingChina Beijing Institute of Biotechnology BeijingChina

Background:The association between 1,3-β-D-glucan(BDG)levels and in-fections caused by pseudomonas aeruginosa or Streptococcus pneumoniae,and the stability of BDG under different storage conditions are unclear.Methods:Strains of pseudomonas aeruginosa and S.pneumoniae were grown in medium and human serum.The BDG concentrations in culture superna-tants were measured.The specificity and stability of BDG were also evaluated.Results:P.aeruginosa produced high levels of BDG in Luria-Bertani medium(>4×10^(4)pg/mL)and human serum(527.0 pg/mL),whereas S.pneumoniae produced low levels of BDG in THY medium(175.6 pg/mL)and human serum(78.3 pg/mL).The BDG produced by these two bacteria was specifically degraded by 1,3-β-D-glucanase.BDG was degraded when stored at different temperatures,decreasing by 22.5%and 9.3%at−20℃and−70℃,respectively,for 63 days;by 30.7%at 4℃for 12 days;and by 12.6%and 22.0%at 37℃for 6 and 12 h.Conclusion:BDG false-positivity must be considered in patients with bacteremia caused by P.aeruginosa when diagnosing invasive fungal infection.Human serum samples for the BDG test in medical facilities should be tested as soon as possible or stored at low temperatures before testing.

关键词： 1,3‐β‐D‐glucan bacteremia false‐positivity pseudomonas aeruginosa stability Streptococcus pneumoniae

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Molecular investigation of exoU and exoY virulence genes in pseudomonas aeruginosa collected from hospitalized patients in North of Iran:A descriptive-analytical study

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Journal of Acute Disease 2024年第2期13卷 74-80页

作者： Ahmad Reza Moradi Mehrdad Gholami Lotfollah Davoodi Negar Hajilou Hamid Reza Goli Sana Institute of Higher Education SariIran Department of Medical Microbiology and Virology Faculty of MedicineMazandaran University of Medical SciencesSariIran Antimicrobial Resistance Research Center Faculty of MedicineMazandaran University of Medical SciencesSariIran Molecular and Cell Biology Research Center Faculty of MedicineMazandaran University of Medical SciencesSariIran

Objective:To investigate the frequency of exoU and exoY genes in patients with pseudomonas aeruginosa infection.Methods:In this study,100 clinical isolates of pseudomonas aeruginosa were collected from patients hospitalized in educational-therapeutic hospitals and were identified using standard microbiological tests.Then,the antibiotic resistance pattern of the isolates was determined by the disk agar diffusion method.The bacterial DNAs were extracted by the alkaline lysis method.Finally,the presence of exoU and exoY genes was evaluated by the PCR test.Results:In this study,47%,72%,29%,39%,40%,and 44%of the isolates were non-susceptible to piperacillin,aztreonam,ceftazidime,imipenem,tobramycin,and ciprofloxacin,respectively.In addition,95%and 93%of the clinical isolates carried the exoU and exoY genes.Blood and fecal isolates had both virulence genes,while only one wound isolate had neither genes.Meanwhile,all urinary isolates contained the exoY gene and only one isolate lacked the exoU gene.Also,88 isolates simultaneously had both exoU and exoY genes.Conclusions:High prevalence of exoU and exoY genes in this region indicates a significant role of typeⅢsecretion system in pathogenesis of pseudomonas aeruginosa.The typeⅢsecretion system may be a suitable target to reduce the pathogenicity of this bacterium.

关键词： pseudomonas aeruginosa exoU exoY Virulence gene TypeⅢsecretion system PCR

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Microbiologically Influenced Corrosion Behavior of Fe_(40)(CoCrMnNi)_(60) and Fe_(60)(CoCrMnNi)_(40) Medium Entropy Alloys in the Presence of pseudomonas aeruginosa

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Acta Metallurgica Sinica(English Letters) 2023年第3期36卷 379-390页

作者： Weiwei Chang Yangyang Li Huaibei Zheng Hongchang Qian Dawei Guo Shuyuan Zhang Yuntian Lou Chi Tat Kwok Lap Mou Tam Dawei Zhang National Materials Corrosion and Protection Data Center Institute for Advanced Materials and TechnologyUniversity of Science and Technology BeijingBeijing 100083China BRI Southeast Asia Network for Corrosion and Protection(MOE) Shunde Graduate School of University of Science and Technology BeijingFoshan 528399China State Key Laboratory of Metal Material for Marine Equipment and Application Anshan 114002China Department of Electromechanical Engineering University of MacaoMacao 999078China Institute for the Development and Quality Macao 999078China

In this work,the microbiologically influenced corrosion(MIC)of Fe_(40)(CoCrMnNi)_(60) and Fe_(60)(CoCrMnNi)_(40) medium entropy alloys(MEAs)induced by pseudomonas aeruginosa(P.aeruginosa)was investigated.Corrosion behaviors during 14 days of immersion in sterile and P.aeruginosa-inoculated culture media are presented.Under sterile conditions,both MEAs exhibited good corrosion resistance against the culture medium solution.In the presence of P.aeruginosa,the pitting corrosion of MEAs was promoted.The results of inductively coupled plasma‒mass spectrometry(ICP‒MS)and potentiodynamic polarization tests showed that the presence of P.aeruginosa promoted the selective dissolution of passive film and accelerated the corrosion of MEAs.The results of X-ray photoelectron spectroscopy(XPS)and Mott-Schottky measurements further demonstrated the degradation effect of P.aeruginosa on the passive film.Compared with Fe_(60)(CoCrMnNi)_(40),Fe_(40)(CoCrMnNi)_(60) manifested better resistance to the MIC caused by P.aeruginosa,which may be attributed to more Cr oxides and fewer Fe oxides of the passive film.

关键词： Microbiologically influenced corrosion Medium entropy alloys pseudomonas aeruginosa

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Nobachelins,new siderophores from Nocardiopsis baichengensis protecting Caenorhabditis elegans from pseudomonas aeruginosa infection

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Synthetic and Systems Biotechnology 2023年第4期8卷 640-646页

作者： Haowen Zhao Yuhao Ren Feng Xie Huanqin Dai Hongwei Liu Chengzhang Fu Rolf Müller Helmholtz Institute for Pharmaceutical Research Saarland(HIPS) Helmholtz Centre for Infection Research(HZI)and Department of PharmacySaarland University66123SaarbrückenGermany Helmholtz International Lab for Anti-Infectives Helmholtz Center for Infection Research38124BraunschweigGermany Institute of Marine Biology and Pharmacology Ocean CollegeZhejiang University316021ZhoushanChina State Key Laboratory of Drug Research Shanghai Institute of Materia MedicaChinese Academy of Sciences555 Zuchongzhi Road201203ShanghaiChina State Key Lab of Mycology Institute of MicrobiologyChinese Academy of Sciences100101BeijingChina

The biosynthetic potential of actinobacteria to produce novel natural products is still regarded as immense.In this paper,we correlated a cryptic biosynthetic gene cluster to chemical molecules by genome mining and chemical analyses,leading to the discovery of a new group of catecholate-hydroxamate siderophores,nobachelins,from Nocardiopsis baichengensis DSM 44845.Nobachelin biosynthesis genes are conserved in several bacteria from the family Nocardiopsidaceae.Structurally,nobachelins feature fatty-acylated hydroxy-ornithine and a rare chlorinated catecholate group.Intriguingly,nobachelins rescued Caenorhabditis elegans from pseudomonas aeruginosa-mediated killing.

关键词： Nocardiopsis Genome mining Siderophore pseudomonas aeruginosa Infection

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Cell division factor ZapE regulates pseudomonas aeruginosa biofilm formation by impacting the pqs quorum sensing system

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mLife 2023年第1期2卷 28-42页

作者： Xi Liu Minlu Jia Jing Wang Hang Cheng Zhao Cai Zhaoxiao Yu Yang Liu Luyan ZMa Lianhui Zhang Yingdan Zhang Liang Yang State Key Laboratory of Microbial Resources Institute of MicrobiologyChinese Academy of SciencesBeijingChina Key University Laboratory of Metabolism and Health of Guangdong School of MedicineSouthern University of Science and TechnologyShenzhenChina Guangdong Province Key Laboratory of Microbial Signals and Disease Control Integrative Microbiology Research CenterSouth China Agricultural UniversityGuangzhouChina Medical Research Center Southern University of Science and Technology HospitalShenzhenChina Shenzhen Third People's Hospital National Clinical Research Center for Infectious DiseaseThe Second Affiliated Hospital of Southern University of Science and TechnologyShenzhenChina

pseudomonas aeruginosa is one of the leading nosocomial pathogens that causes both severe acute and chronic infections.The strong capacity of P.aeruginosa to form biofilms can dramatically increase its antibiotic resistance and lead to treatment failure.The biofilm resident bacterial cells display distinct gene expression profiles and phenotypes compared to their free-living counterparts.Elucidating the genetic determinants of biofilm formation is crucial for the development of antibiofilm drugs.In this study,a highthroughput transposon-insertion site sequencing(Tn-seq)approach was employed to identify novel P.aeruginosa biofilm genetic determinants.When analyzing the novel biofilm regulatory genes,we found that the cell division factor ZapE(PA4438)controls the P.aeruginosa pqs quorum sensing system.The ΔzapE mutant lost fitness against the wild-type PAO1 strain in biofilms and its production of 2-heptyl-3-hydroxy-4(1H)-quinolone(PQS)had been reduced.Further biochemical analysis showed that ZapE interacts with PqsH,which encodes the synthase that converts 2-heptyl-4-quinolone(HHQ)to PQS.In addition,site-directed mutagenesis of the ATPase active site of ZapE(K72A)abolished the positive regulation of ZapE on PQS signaling.As ZapE is highly conserved among the pseudomonas group,our study suggests that it is a potential drug target for the control of pseudomonas infections.

关键词： biofilm pseudomonas aeruginosa pseudomonas quinolone signal transposon‐insertion site sequencing(Tn‐seq) ZapE

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The transcriptional regulators of virulence for pseudomonas aeruginosa:Therapeutic opportunity and preventive potential of its clinical infections

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Genes & Diseases 2023年第5期10卷 2049-2063页

作者： Xiaolong Shao Chunyan Yao Yiqing Ding Haiyan Hu Guoliang Qian Mingliang He Xin Deng College of Plant Protection Laboratory of Plant ImmunityKey Laboratory of Integrated Management of Crop Diseases and PestsNanjing Agricultural UniversityNanjingJiangsu 210095China Department of Biomedical Sciences City University of Hong KongKowloon TongHong Kong SARChina School of Pharmaceutical Sciences Sun Yat-Sen UniversityUniversity TownGuangzhouGuangdong 510006China Tung Biomedical Sciences Centre City University of Hong KongKowloon TongHong Kong SARChina Shenzhen Research Institute City University of Hong KongShenzhenGuangdong 518057China

In pseudomonas aeruginosa(P.aeruginosa),transcription factors(TFs)are important mediators in the genetic regulation of adaptability and pathogenicity to respond to multiple environmental stresses and host defences.The P.aeruginosa genome harbours 371 putative TFs;of these,about 70 have been shown to regulate virulence-associated phenotypes by binding to the promoters of their target genes.Over the past three decades,several techniques have been applied to identify TF binding sites on the P.aeruginosa genome,and an atlas of TF binding patterns has been mapped.The virulence-associated regulons of TFs show complex crosstalk in P.aeruginosa's regulatory network.In this review,we summarise the recent literature on TF regulatory networks involved in the quorum-sensing system,biofilm formation,pyocyanin synthesis,motility,the type III secretion system,the type VI secretion system,and oxidative stress responses.We discuss future perspectives that could provide insights and targets for preventing clinical infections caused by P.aeruginosa based on the global regulatory network of transcriptional regulators.

关键词： Crosstalk pseudomonas aeruginosa Regulatory network Transcriptional regulators Virulence

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β-cyclopiazonic acid binds iron demonstrating siderophorelike activity and promotes growth in pseudomonas aeruginosa

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Journal of Oceanology and Limnology 2023年第3期41卷 1159-1167页

作者： Yuqi GUO Youmin YING Qihao WU Bin WEI Jianwei CHEN Hong WANG College of Pharmaceutical Science&Collaborative Innovation Center of Yangtze River Delta Region Green Pharmaceuticals Key Laboratory of Marine Fishery Resources Exploitment&Utilization of Zhejiang ProvinceZhejiang University of TechnologyHangzhou 310014China

This chemical study reports a novel siderophore-like compound,β-cyclopiazonic acid(1,β-CPA)extracted from marine fungus Aspergillus flavus.The chemical structure ofβ-CPA was elucidated by a combination of extensive spectroscopic analyses and TDDFT-ECD calculations.The iron-binding ability and CAS assays demonstrate thatβ-CPA is a novel siderophore that features a different chemical structure from those of traditional siderophores.Theβ-CPA has no obvious influence on the growth of bacterium pseudomonas aeruginosa PAO1.However,its iron chelator could promote the growth of P.aeruginosa PAO1,suggesting that P.aeruginosa employed siderophores to sequester iron,which is vital for their survival.The study provides the physiochemical evaluation ofβ-CPA,an unusual skeletonstructure siderophore,which for the first time,was proven to have the ability to bind iron and affect P.aeruginosa growth.This new discovery of siderophore provides an opportunity for developing novel anti-P.aeruginosa drugs.

关键词： marine fungi β-cyclopiazonic acid siderophore pseudomonas aeruginosa

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Infl uence of nitrate concentrations on EH40 steel corrosion aff ected by coexistence of Desulfovibrio desulfuricans and pseudomonas aeruginosa bacteria

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Journal of Oceanology and Limnology 2022年第4期40卷 1448-1461页

作者： Zhihua SUN Jiajia WU Dun ZHANG Ce LI Liyang ZHU Ee LI Key Laboratory of Marine Environmental Corrosion and Biofouling Institute of OceanologyChinese Academy of SciencesQingdao 266071China Open Studio for Marine Corrosion and Protection Pilot National Laboratory for Marine Science and Technology(Qingdao)Qingdao 266237China Center for Ocean Mega-Science Chinese Academic of SciencesQingdao 266071China University of Chinese Academy of Sciences Beijing 100049China Key Laboratory of Marine Materials and Related Technologies Ningbo Institute of Materials Technology&EngineeringChinese Academy of SciencesNingbo 315201China

Nitrate addition is a common bio-competitive exclusion(BCE)method to mitigate corrosion in produced water reinjection systems,which can aff ect microbial community compositions,especially nitrate and sulfate reducing bacteria,but its eff ectiveness is in controversy.We investigated the infl uence of nitrate concentrations on EH40 steel corrosion aff ected by coexistence of Desulfovibrio vulgaris and pseudomonas aeruginosa bacteria.Results demonstrate that only mixed bacteria or nitrate had little eff ect on EH40 steel corrosion,and nitrate could accelerate the corrosion of EH40 steel through the action of microorganisms.The corrosion promotion of nitrate was dependent on its concentrations,which increased from 0 to 5 g/L and decreased from 5 to 50 g/L.These diff erences were believed to be related to the regulation of nitrate in the growth of bacteria and biofi lms.Therefore,care must be taken to BCE method with nitrate when nitrate reducing bacteria with high corrosive activity are present in the environments.

关键词： microbiologically influenced corrosion nitrate Desulfovibrio vulgaris pseudomonas aeruginosa

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