المؤلفون: Zhenwei Xia, Zhihan Wang, Shugang Qin, Ping Lin, Pan Gao, Min Wu, Nadeem Khan, Colin K. Combs, Kai Guo, Qinqin Pu

المصدر: Signal Transduction and Targeted Therapy, Vol 6, Iss 1, Pp 1-10 (2021)
Signal Transduction and Targeted Therapy

مصطلحات موضوعية: 0301 basic medicine, Cancer Research, Peroxisome proliferator-activated receptor gamma, Cytoplasm, Neutrophils, QH301-705.5, Inflammation, medicine.disease_cause, Perilipin-2, Article, Receptors, G-Protein-Coupled, 03 medical and health sciences, Mice, 0302 clinical medicine, 4-Butyrolactone, Tongue, Taste receptor, Lipid droplet, Genetics, medicine, Homoserine, Animals, Humans, Pseudomonas Infections, Biology (General), Receptor, Cell Nucleus, Chemistry, Pseudomonas aeruginosa, Lipid Droplets, Peroxisome, Cell biology, PPAR gamma, 030104 developmental biology, Host-Pathogen Interactions, Infectious diseases, Medicine, medicine.symptom, Infection, 030217 neurology & neurosurgery

الوصف: Bitter receptors function primarily in sensing taste, but may also have other functions, such as detecting pathogenic organisms due to their agile response to foreign objects. The mouse taste receptor type-2 member 138 (TAS2R138) is a member of the G-protein-coupled bitter receptor family, which is not only found in the tongue and nasal cavity, but also widely distributed in other organs, such as the respiratory tract, gut, and lungs. Despite its diverse functions, the role of TAS2R138 in host defense against bacterial infection is largely unknown. Here, we show that TAS2R138 facilitates the degradation of lipid droplets (LDs) in neutrophils during Pseudomonas aeruginosa infection through competitive binding with PPARG (peroxisome proliferator-activated receptor gamma) antagonist: N-(3-oxododecanoyl)-l-homoserine lactone (AHL-12), which coincidently is a virulence-bound signal produced by this bacterium (P. aeruginosa). The released PPARG then migrates from nuclei to the cytoplasm to accelerate the degradation of LDs by binding PLIN2 (perilipin-2). Subsequently, the TAS2R138–AHL-12 complex targets LDs to augment their degradation, and thereby facilitating the clearance of AHL-12 in neutrophils to maintain homeostasis in the local environment. These findings reveal a crucial role for TAS2R138 in neutrophil-mediated host immunity against P. aeruginosa infection.

الوصول الحر: https://explore.openaire.eu/search/publication?articleId=doi_dedup___::28d498d93007663dc6980d9801a46369Test
https://doaj.org/article/6d22490f6a5946e390f336d307620035Test

المؤلفون: Ping Lin, Guanwang Shen, Kai Guo, Shugang Qin, Qinqin Pu, Zhihan Wang, Pan Gao, Zhenwei Xia, Nadeem Khan, Jianxin Jiang, Qingyou Xia, Min Wu

المصدر: Nucleic acids research. 50(8)

مصطلحات موضوعية: Mice, SARS-CoV-2, viruses, Genetics, virus diseases, Animals, COVID-19, Humans, RNA Editing, Antiviral Agents, Pandemics, respiratory tract diseases, RNA, Guide, Kinetoplastida

الوصف: Gene-editing technologies, including the widespread usage of CRISPR endonucleases, have the potential for clinical treatments of various human diseases. Due to the rapid mutations of SARS-CoV-2, specific and effective prevention and treatment by CRISPR toolkits for coronavirus disease 2019 (COVID-19) are urgently needed to control the current pandemic spread. Here, we designed Type III CRISPR endonuclease antivirals for coronaviruses (TEAR-CoV) as a therapeutic to combat SARS-CoV-2 infection. We provided a proof of principle demonstration that TEAR-CoV-based RNA engineering approach leads to RNA-guided transcript degradation both in vitro and in eukaryotic cells, which could be used to broadly target RNA viruses. We report that TEAR-CoV not only cleaves SARS-CoV-2 genome and mRNA transcripts, but also degrades live influenza A virus (IAV), impeding viral replication in cells and in mice. Moreover, bioinformatics screening of gRNAs along RNA sequences reveals that a group of five gRNAs (hCoV-gRNAs) could potentially target 99.98% of human coronaviruses. TEAR-CoV also exerted specific targeting and cleavage of common human coronaviruses. The fast design and broad targeting of TEAR-CoV may represent a versatile antiviral approach for SARS-CoV-2 or potentially other emerging human coronaviruses.

الوصول الحر: https://explore.openaire.eu/search/publication?articleId=doi_dedup___::d4044aa8973456ef0048b9d6dee7f77fTest
https://pubmed.ncbi.nlm.nih.gov/35166837Test

المؤلفون: Cong Chen, Sidong Cai, Min Wu, Run Wang, Mingya Liu, Gaozhen Cao, Min Dong, Kai-Hang Yiu

المصدر: Disease Markers.

مصطلحات موضوعية: Inflammation, TNF Receptor-Associated Factor 6, Article Subject, Macrophages, Biochemistry (medical), Clinical Biochemistry, Myocardial Infarction, General Medicine, Macrophage Activation, Exosomes, Mice, MicroRNAs, Genetics, Animals, Myocytes, Cardiac, Molecular Biology

الوصف: Myocardial infarction arises from an excessive or prolonged inflammatory response, leading to ventricular remodeling or impaired cardiac function. Macrophages exhibit different polarization types associated with inflammation both at steady state and after myocardial infarction. Exosomal miR-146a-5p has been identified as an important molecule in the cardiovascular field in recent years. However, the effect of cardiomyocyte-derived exosomal miR-146a-5p on macrophages has not yet been elucidated. Initially, we found that exosomes with low expression of miR-146a-5p derived from myocardial infarction tissues modulated macrophage polarization. To determine whether cardiomyocyte-derived exosomal miR-146a-5p mediated macrophage polarization, we treated macrophages with exosomes rich in miR-146a-5p collected from neonatal mouse cardiomyocytes. The effects of exosomal miR-146a-5p on macrophage polarization were measured using RT-qPCR, transwell assays, and western blotting. The results showed that the increased expression of miR-146a-5p promoted M1 macrophage polarization, inhibited M2 macrophage polarization, and increased the expression of VEGFA. However, the decreased expression of exosomalmiR-146a-5p showed the opposite trends. Interestingly, in contrast to treatment with the solitary miR-146a-5p mimic, exosomal miR-146a-5p derived from neonatal mouse cardiomyocytes reduced TNFα and iNOS expression. In addition, when macrophages were activated by the miR-146a-5p mimic or exosomal miR-146a-5p, the expression of TNF receptor-associated factor 6 (TRAF6), a target gene of miR-146a-5p, was reduced significantly. Taken together, these findings indicate that exosomal miR-146a-5p derived from cardiomyocytes could stimulate M1 macrophage polarization to induce an inflammatory reaction, while targeting TRAF6, exerting an anti-inflammatory effect. Exosomal miR-146a-5p plays important roles in macrophages, illuminating a novel potential therapeutic target in myocardial infarction.

وصف الملف: text/xhtml

الوصول الحر: https://explore.openaire.eu/search/publication?articleId=doi_dedup___::ce07ddfe2fc91aef5c6b38471f5fe39fTest

المؤلفون: Meng Chen, Yu-Juan Mao, Qiu Zhao, Ping Chen, Mei Ye, Mingxing Zhuo, Hua Zhu, Lian-Yun Li, Ya-Li Yu, Min Wu

المصدر: Clinical Epigenetics

مصطلحات موضوعية: China, IBD, Biology, Inflammatory bowel disease, Epigenesis, Genetic, Pathogenesis, Mice, STAT1, Genetics, medicine, Animals, Humans, Colitis, Enhancer, EP300, Molecular Biology, Transcription factor, Genetics (clinical), Acute colitis, Adaptor Proteins, Signal Transducing, Research, DNA Methylation, Inflammatory Bowel Diseases, Phosphoproteins, H3K27ac, medicine.disease, Disease Models, Animal, STAT1 Transcription Factor, biology.protein, Cancer research, Cytokines, E1A-Associated p300 Protein, Developmental Biology

الوصف: Background The aetiology of inflammatory bowel disease (IBD) is related to genetics and epigenetics. Epigenetic regulation of the pathogenesis of IBD has not been well defined. Here, we investigated the role of H3K27ac events in the pathogenesis of IBD. Based on previous ChIP-seq and RNA-seq assays, we studied signal transducer and activator of transcription 1 (STAT1) as a transcription factor (TF) and investigated whether the STAT1–EP300–H3K27ac axis contributes to the development of IBD. We performed ChIP-PCR to investigate the interaction between STAT1 and H3K27ac, and co-IP assays were performed to investigate the crosstalk between STAT1 and EP300. Results Lymphocyte cytosolic protein 2 (LCP2) and TNF-α‐inducible protein 2 (TNFAIP2) are target genes of STAT1. p-STAT1 binds to the enhancer loci of the two genes where H3K27ac is enriched, and EP300 subsequently binds to regulate their expression. In mice with dextran sulfate sodium (DSS)-induced acute colitis, an EP300 inhibitor significantly inhibited colitis. Conclusions p-STAT1 and EP300 promote TNFAIP2 and LCP2 expression through an increase in H3K27ac enrichment on their enhancers and contribute to the pathogenesis of chronic inflammation. Graphic abstract

الوصول الحر: https://explore.openaire.eu/search/publication?articleId=doi_dedup___::7c2064c54113b415f0ec500f42c6fcbcTest
https://doi.org/10.1186/s13148-021-01101-wTest

المؤلفون: Min Wu, Li Zhang, Zhenling Wang

المصدر: Signal Transduction and Targeted Therapy
Signal Transduction and Targeted Therapy, Vol 5, Iss 1, Pp 1-3 (2020)

مصطلحات موضوعية: RNA Caps, Cancer Research, RNA splicing, Transcription, Genetic, Recombinant Fusion Proteins, Mutant Chimeric Proteins, Virulence, lcsh:Medicine, Biology, Virus Replication, Cell Line, Chimera (genetics), Mice, Open Reading Frames, Viral Proteins, Dogs, RNA Virus Infections, Cricetinae, Genetics, T cell immunity, Animals, Humans, RNA Viruses, RNA, Messenger, lcsh:QH301-705.5, Novel protein, lcsh:R, RNA-Dependent RNA Polymerase, Virology, Research Highlight, lcsh:Biology (General), Influenza A virus, RNA, Viral, Cattle, Infection, 5' Untranslated Regions

الوصف: RNA viruses are a major human health threat. The life cycles of many highly pathogenic RNA viruses like influenza A virus (IAV) and Lassa virus depends on host mRNA, because viral polymerases cleave 5'-m7G-capped host transcripts to prime viral mRNA synthesis ("cap-snatching"). We hypothesized that start codons within cap-snatched host transcripts could generate chimeric human-viral mRNAs with coding potential. We report the existence of this mechanism of gene origination, which we named "start-snatching." Depending on the reading frame, start-snatching allows the translation of host and viral "untranslated regions" (UTRs) to create N-terminally extended viral proteins or entirely novel polypeptides by genetic overprinting. We show that both types of chimeric proteins are made in IAV-infected cells, generate T cell responses, and contribute to virulence. Our results indicate that during infection with IAV, and likely a multitude of other human, animal and plant viruses, a host-dependent mechanism allows the genesis of hybrid genes.

الوصول الحر: https://explore.openaire.eu/search/publication?articleId=doi_dedup___::08d7ddb0d8246b9c949379c3e34ff4d2Test
http://europepmc.org/articles/PMC7445444Test

المؤلفون: Zhiyao He, Xiangxian Zhang, Li Liu, Yan Tie, Xiawei Wei, Min Wu, Yu Liu, Hongyi Li, Min Luo, Bin Shao, Jingyun Yang, Xia Yuan, Heng Zheng

المصدر: Signal Transduction and Targeted Therapy
Signal Transduction and Targeted Therapy, Vol 5, Iss 1, Pp 1-15 (2020)

مصطلحات موضوعية: 0301 basic medicine, Cancer Research, Lung Neoplasms, Angiogenesis, medicine.medical_treatment, lcsh:Medicine, Apoptosis, Article, Metastasis, Targeted therapy, Mice, 03 medical and health sciences, Prognostic markers, Folic Acid, 0302 clinical medicine, Tumor-Associated Macrophages, Tumor Microenvironment, Genetics, medicine, Animals, Humans, Folate Receptor 2, Lung cancer, lcsh:QH301-705.5, Cell Proliferation, Tumor microenvironment, business.industry, lcsh:R, Cancer, medicine.disease, 030104 developmental biology, lcsh:Biology (General), A549 Cells, Folate receptor, 030220 oncology & carcinogenesis, Liposomes, Cancer research, lipids (amino acids, peptides, and proteins), business

الوصف: Tumor-associated macrophages (TAMs) facilitate cancer progression by promoting tumor invasion, angiogenesis, metastasis, inflammatory responses, and immunosuppression. Folate receptor β (FRβ) is overexpressed in TAMs. However, the clinical significance of FRβ-positive macrophages in lung cancer remains poorly understood. In this study, we verified that FRβ overexpression in lung cancer TAMs was associated with poor prognosis. We utilized a folate-modified lipoplex comprising a folate-modified liposome (F-PLP) delivering a BIM-S plasmid to target both lung cancer cells and FRβ-positive macrophages in the tumor microenvironment. Transfection of LL/2 cells and MH-S cells with F-PLP/pBIM induced cell apoptosis. Injection of F-PLP/pBIM into LL/2 and A549 lung cancer models significantly depleted FRβ-positive macrophages and reduced tumor growth. Treatment of tumor-bearing mice with F-PLP/pBIM significantly inhibited tumor growth in vivo by inducing tumor cell and macrophage apoptosis, reducing tumor proliferation, and inhibiting tumor angiogenesis. In addition, a preliminary safety evaluation demonstrated a good safety profile of F-PLP/pBIM as a gene therapy administered intravenously. This work describes a novel application of lipoplexes in lung cancer targeted therapy that influences the tumor microenvironment by targeting TAMs.

الوصول الحر: https://explore.openaire.eu/search/publication?articleId=doi_dedup___::4721efc600e486cab95836dcb03c2af1Test
http://europepmc.org/articles/PMC6976681Test

المؤلفون: Yuan Zhu, Qi-Xin Hu, Hui-Yi Wang, Lingao Ju, Lu Jiang, Bo Zhong, Min Wu, Zhen Wang, Chen-Yu Wang, Lian-Yun Li

المصدر: PLoS Pathogens, Vol 17, Iss 9, p e1009918 (2021)
PLoS Pathogens

مصطلحات موضوعية: RNA viruses, viruses, Regulator, Pathology and Laboratory Medicine, Biochemistry, Ligases, Mice, RNA Virus Infections, Chlorocebus aethiops, Demethylase activity, Medicine and Health Sciences, Post-Translational Modification, Amino Acids, Biology (General), Histone Demethylases, biology, Organic Compounds, RIG-I, Chemistry, virus diseases, Small interfering RNA, Enzymes, Precipitation Techniques, Cell biology, Nucleic acids, Histone, Vesicular Stomatitis Virus, Medical Microbiology, Viral Pathogens, Viruses, Physical Sciences, Pathogens, Basic Amino Acids, Signal transduction, Research Article, TRIM25, animal structures, QH301-705.5, Immunoblotting, Immunology, Molecular Probe Techniques, Receptors, Cell Surface, chemical and pharmacologic phenomena, Transfection, Research and Analysis Methods, Microbiology, Rhabdoviruses, Virology, Genetics, Animals, Humans, Immunoprecipitation, Molecular Biology Techniques, Non-coding RNA, Vero Cells, Microbial Pathogens, Molecular Biology, Lysine, Organic Chemistry, Ubiquitination, Organisms, Chemical Compounds, Biology and Life Sciences, Proteins, RC581-607, Co-Immunoprecipitation, Gene regulation, Mice, Inbred C57BL, HEK293 Cells, Gene Expression Regulation, Viral replication, Enzymology, biology.protein, RNA, Demethylase, Parasitology, Gene expression, Immunologic diseases. Allergy

الوصف: Under RNA virus infection, retinoic acid-inducible gene I (RIG-I) in host cells recognizes viral RNA and activates the expression of type I IFN. To investigate the roles of protein methyltransferases and demethylases in RIG-I antiviral signaling pathway, we screened all the known related enzymes with a siRNA library and identified LSD1 as a positive regulator for RIG-I signaling. Exogenous expression of LSD1 enhances RIG-I signaling activated by virus stimulation, whereas its deficiency restricts it. LSD1 interacts with RIG-I, promotes its K63-linked polyubiquitination and interaction with VISA/MAVS. Interestingly, LSD1 exerts its function in antiviral response not dependent on its demethylase activity but through enhancing the interaction between RIG-I with E3 ligases, especially TRIM25. Furthermore, we provide in vivo evidence that LSD1 increases antiviral gene expression and inhibits viral replication. Taken together, our findings demonstrate that LSD1 is a positive regulator of signaling pathway triggered by RNA-virus through mediating RIG-I polyubiquitination.
Author summary RIG-I signaling pathway is critical for human cells to defend from RNA virus infection, such as SARS-CoV-2, influenza virus, and Vesicular Stomatitis Virus (VSV). LSD1 is a histone demethylase regulating transcription. The current study reveals a novel function of LSD1 in regulating the activation of RIG-I signaling pathway. LSD1 interacts with RIG-I and promotes RIG-I poly-ubiquitination independent of its demethylase activity. LSD1 facilitates the interaction between RIG-I and its ubiquitin E3 ligase TRIM25, which is crucial for recruitment of downstream proteins. The mice with LSD1 deficiency are susceptible to virus infection and have lower survival rate. Taken together, our findings demonstrate a novel molecular mechanism for regulating the anti-viral RIG-I signaling pathway.

الوصول الحر: https://explore.openaire.eu/search/publication?articleId=doi_dedup___::3d2b77951e6a4d3bff8618097d50e8b2Test
https://doi.org/10.1371/journal.ppat.1009918Test

المؤلفون: Quan-Min Wu, Jiang Li, Xiao-Qing Wang, Cheng-Qiang Zhang

المصدر: DNA and Cell Biology. 36:1117-1125

مصطلحات موضوعية: Male, 0301 basic medicine, Epithelial-Mesenchymal Transition, Mice, Nude, Bone Neoplasms, Vimentin, Bioinformatics, medicine.disease_cause, Metastasis, Mice, 03 medical and health sciences, 0302 clinical medicine, Cell Line, Tumor, microRNA, Genetics, medicine, Animals, Humans, Gene silencing, Neoplasm Invasiveness, Neoplasm Metastasis, 3' Untranslated Regions, Molecular Biology, Cell Proliferation, Mice, Inbred BALB C, Osteosarcoma, Gene knockdown, biology, Cell Biology, General Medicine, Prognosis, medicine.disease, Long non-coding RNA, Gene Expression Regulation, Neoplastic, MicroRNAs, 030104 developmental biology, Gene Knockdown Techniques, 030220 oncology & carcinogenesis, Cancer research, biology.protein, RNA, Long Noncoding, Carcinogenesis

الوصف: Long noncoding RNAs (lncRNAs) have been illustrated to function as important regulator in carcinogenesis and cancer progression. However, roles of lncRNA miR210HG (miR210 host gene) in osteosarcoma remain unclear. In this study, miR210HG expression level was significantly upregulated in 55 cases of osteosarcoma tissue samples compared to adjacent normal tissue. Besides, the aberrantly enhanced miR210HG expression predicted poor prognosis and lower survival rate. In vitro, miR210HG knockdown suppressed the osteosarcoma cell proliferation, invasion, and epithelial-mesenchymal transition-related marker (N-cadherin and vimentin) expression. In vivo, miR210HG silencing decreased the tumor growth. miR-503 was verified to be the target miRNA of miR210HG using bioinformatics online program and luciferase assay. Furthermore, miR-503 could reverse the role of miR210HG on osteosarcoma cells. In conclusion, our study indicates that miR210HG sponges miR-503 to facilitate osteosarcoma cell invasion and metastasis, revealing the oncogenic role of miR210HG on osteosarcoma cells.

الوصول الحر: https://explore.openaire.eu/search/publication?articleId=doi_dedup___::d1f187185d89cf008db1256a8048b00aTest
https://doi.org/10.1089/dna.2017.3888Test

المؤلفون: Pei‑Hui Ding, Lili Chen, Xiao‑Tao Chen, Jing‑Yi Tan, Li‑Hong Lei, Yan‑Min Wu

المصدر: Molecular Medicine Reports

مصطلحات موضوعية: 0301 basic medicine, RAC-β serine/threonine protein kinase, Cancer Research, Cell, Osteocalcin, Core Binding Factor Alpha 1 Subunit, Biology, Biochemistry, interleukin-17A, 03 medical and health sciences, Mice, Phosphatidylinositol 3-Kinases, 0302 clinical medicine, Calcification, Physiologic, Genetics, medicine, Animals, Humans, Periodontitis, Molecular Biology, PI3K/AKT/mTOR pathway, Cell Proliferation, Osteoblasts, Cell growth, osteoblast calcification, Interleukin-17, Osteoblast, Cell Differentiation, 030206 dentistry, Articles, Cell cycle, medicine.disease, Alkaline Phosphatase, Cell biology, 030104 developmental biology, medicine.anatomical_structure, Oncology, Gene Expression Regulation, osteoblast differentiation, Gene Knockdown Techniques, Molecular Medicine, Alkaline phosphatase, Signal transduction, Proto-Oncogene Proteins c-akt, Calcification, Signal Transduction

الوصف: Interleukin (IL)‑17A exhibits pleiotropic biological activities and serves a role in the progression of periodontitis. However, data describing the association between IL‑17 and osteogenesis are not conclusive. It was previously demonstrated that RAC‑β serine/threonine protein kinase (AKT2)‑specific knockdown in MC3T3‑E1 cells weakened osteogenic effects. The role of AKT2 in the regulation of IL‑17A for osteoblast differentiation and calcification remains unclear. The MTT method was adopted in the present study to assess cell proliferation; cell cycle distribution was analyzed by flow cytometry. Following osteogenic induction treatment, the involvement of phosphatidylinositol 3‑kinase (PI3K) and phosphorylated‑PI3K was evaluated by western blotting. The effects of IL‑17A on osteogenesis‑associated markers, including Runt‑related transcription factor 2 (Runx‑2), alkaline phosphatase (ALP) and osteocalcin (OCN) were evaluated by reverse transcription‑quantitative polymerase chain reaction (RT‑qPCR) analysis. An ALP activity assay and Alizarin Red S staining were used to assess the differentiation and calcification functions. AKT2 knockdown inhibited MC3T3‑E1 cell proliferation, inducing significantly increased G0/G1 cell counts, and reduced S and G2/M cell numbers. IL‑17A exerted no significant effects. The protein levels of p‑PI3K, gene expression levels of IL‑17A, Runx‑2, ALP and OCN, and relative ALP activity and calcification areas were increased in the induction group, and these effects were markedly promoted by treatment with IL‑17A. AKT2 knockdown in MC3T3‑E1 cells resulted in reduced IL‑17A‑induced differentiation and calcification, although it was not completely inhibited. The results of the present study suggested that AKT2 signaling was required for MC3T3‑E1 cell proliferation. IL‑17A promoted osteoblast differentiation and calcification in a partly AKT2‑dependent manner in MC3T3‑E1 cells in vitro, possibly reflecting compensation by other signaling pathways. The results of the present study may offer novel perspectives to guide the clinical strategy for the prevention and treatment of periodontitis.

الوصول الحر: https://explore.openaire.eu/search/publication?articleId=doi_dedup___::ce984650a96b5f1c0414ed4b09b10f61Test
http://europepmc.org/articles/PMC5865781Test

المؤلفون: Gukui Chen, Tietao Wang, Yani Zhang, Yuying Han, Min Wu, Haihua Liang, Qiong Liu, Qinqin Pu, Linghui Xu

المصدر: PLoS Pathogens
PLoS Pathogens, Vol 15, Iss 12, p e1008198 (2019)

مصطلحات موضوعية: Physiology, Cell Membranes, Secretion Systems, Artificial Gene Amplification and Extension, medicine.disease_cause, Pathology and Laboratory Medicine, Polymerase Chain Reaction, Biochemistry, Mice, Microbial Physiology, Medicine and Health Sciences, Bacterial Physiology, Biology (General), Post-Translational Modification, 0303 health sciences, Virulence, Chemistry, Effector, Chromatographic Techniques, Pseudomonas Aeruginosa, Type VI Secretion Systems, Cell biology, Bacterial Pathogens, DNA-Binding Proteins, Medical Microbiology, Amino Acid Specific Chromatography, Azurin, Pathogens, Cellular Structures and Organelles, Bacterial outer membrane, Signal Peptides, Research Article, Signal peptide, QH301-705.5, Virulence Factors, Immunology, DNA construction, Research and Analysis Methods, Microbiology, 03 medical and health sciences, Bacterial Proteins, Virology, Pseudomonas, Genetics, medicine, Glutathione Chromatography, Animals, Secretion, Pseudomonas Infections, Molecular Biology Techniques, Microbial Pathogens, Molecular Biology, 030304 developmental biology, Type VI secretion system, Bacteria, 030306 microbiology, Pseudomonas aeruginosa, Affinity Chromatography, Organisms, Biology and Life Sciences, Membrane Proteins, Proteins, Bacteriology, Cell Biology, RC581-607, Outer Membrane Proteins, Plasmid Construction, Parasitology, Immunologic diseases. Allergy, Physiological Processes, Copper

الوصف: The type VI secretion system (T6SS) is widely distributed in Gram-negative bacteria, whose function is known to translocate substrates to eukaryotic and prokaryotic target cells to cause host damage or as a weapon for interbacterial competition. Pseudomonas aeruginosa encodes three distinct T6SS clusters (H1-, H2-, and H3-T6SS). The H1-T6SS-dependent substrates have been identified and well characterized; however, only limited information is available for the H2- and H3-T6SSs since relatively fewer substrates for them have yet been established. Here, we obtained P. aeruginosa H2-T6SS-dependent secretomes and further characterized the H2-T6SS-dependent copper (Cu2+)-binding effector azurin (Azu). Our data showed that both azu and H2-T6SS were repressed by CueR and were induced by low concentrations of Cu2+. We also identified the Azu-interacting partner OprC, a Cu2+-specific TonB-dependent outer membrane transporter. Similar to H2-T6SS genes and azu, expression of oprC was directly regulated by CueR and was induced by low Cu2+. In addition, the Azu-OprC-mediated Cu2+ transport system is critical for P. aeruginosa cells in bacterial competition and virulence. Our findings provide insights for understanding the diverse functions of T6SSs and the role of metal ions for P. aeruginosa in bacteria-bacteria competition.
Author summary The type VI secretion system (T6SS) is a specific macromolecular protein export apparatus, and widely distributed in Gram-negative bacteria. T6SS plays an important role in anti-bacterial competition or delivers effector proteins to both eukaryotic and prokaryotic cells. In the present study, we performed secretomes analysis and identified 21 substrates of P. aeruginosa H2-T6SS-dependent. Specifically, we report a Cu2+-scavenging pathway consisting of a copper transporter, OprC, and a type VI secretion system (H2-T6SS)-secreted Cu2+-binding protein, Azu. Both of them are under control of the transcriptional regulator CueR. Indeed, the Azu-OprC-mediated Cu2+ transport system is critical for P. aeruginosa cells in bacterial competition and virulence. These findings exemplify how P. aeruginosa deploys this metal system to adapt to the complex environment during evolution.

الوصول الحر: https://explore.openaire.eu/search/publication?articleId=doi_dedup___::a255b6cfcdeb18bd60153c0e6ec5e847Test
https://pubmed.ncbi.nlm.nih.gov/31790504Test