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Construction of high-density transposon mutant library of Staphylococcus aureus using bacteriophage ϕ11
Wonsik Lee
J. Microbiol. 2022;60(12):1123-1129.   Published online November 24, 2022
DOI: https://doi.org/10.1007/s12275-022-2476-2
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AbstractAbstract
Transposon mutant libraries are an important resource to study bacterial metabolism and pathogenesis. The fitness analysis of mutants in the libraries under various growth conditions provides important clues to study the physiology and biogenesis of structural components of a bacterial cell. A transposon library in conjunction with next-generation sequencing techniques, collectively named transposon sequencing (Tnseq), enables high-throughput genome profiling and synthetic lethality analysis. Tn-seq has also been used to identify essential genes and to study the mode of action of antibacterials. To construct a high-density transposon mutant library, an efficient delivery system for transposition in a model bacterium is essential. Here, I describe a detailed protocol for generating a high-density phage-based transposon mutant library in a Staphylococcus aureus strain, and this protocol is readily applicable to other S. aureus strains including USA300 and MW2.

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  • Optimizing phage-based mutant recovery and minimizing heat effect in the construction of transposon libraries in Staphylococcus aureus
    Sally W. Yousief, Nader Abdelmalek, Bianca Paglietti
    Scientific Reports.2024;[Epub]     CrossRef
Lysobacter arenosi sp. nov. and Lysobacter solisilvae sp. nov. isolated from soil
Kyeong Ryeol Kim† , Kyung Hyun Kim† , Shehzad Abid Khan , Hyung Min Kim , Dong Min Han , Che Ok Jeon
J. Microbiol. 2021;59(8):709-718.   Published online June 1, 2021
DOI: https://doi.org/10.1007/s12275-021-1156-y
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AbstractAbstract
Two Gram-stain negative, yellow-pigmented, and mesophilic bacteria, designated strains R7T and R19T, were isolated from sandy and forest soil, South Korea, respectively. Both strains were non-motile rods showing catalase- and oxidase-positive activities. Both strains were shown to grow at 10–37°C and pH 6.0–9.0, and in the presence of 0–1.5% (w/v) NaCl. Strain R7T contained iso-C14:0, iso-C15:0, iso-C16:0, and summed feature 9 (comprising C16:0 10-methyl and/or iso-C17:1 ω9c), whereas strain R19T contained iso-C11:0 3-OH, C16:1 ω7c alcohol, iso-C11:0, iso-C15:0, iso-C16:0, and summed feature 9 (comprising C16:0 10-methyl and/or iso-C17:1 ω9c) as major cellular fatty acids (> 5%). Both strains contained ubiquinone- 8 as the sole isoprenoid quinone and phosphatidylglycerol, phosphatidylethanolamine, and an unidentified phospholipid as the major polar lipids. The DNA G + C contents of strains R7T and R19T calculated from their genomes were 66.9 mol% and 68.9 mol%, respectively. Strains R7T and R19T were most closely related to Lysobacter panacisoli C8-1T and Lysobacter niabensis GH34-4T with 98.7% and 97.8% 16S rRNA sequence similarities, respectively. Phylogenetic analyses based on 16S rRNA gene sequences showed that strains R7T and R19T formed distinct phylogenetic lineages within the genus Lysobacter. Based on phenotypic, chemotaxonomic, and molecular features, strains R7T and R19T represent novel species of the genus Lysobacter, for which the names Lysobacter arenosi sp. nov. and Lysobacter solisilvae sp. nov. are proposed. The type strains of L. arenosi and L. solisilvae are R7T (= KACC 21663T = JCM 34257T) and R19T (= KACC 21767T = JCM 34258T), respectively.

Citations

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  • Luteimonas flava sp. nov. and Aquilutibacter rugosus gen. nov., sp. nov., isolated from freshwater environments in China and re-examining the taxonomic status of genera Luteimonas and Lysobacter
    Huibin Lu, Li Chen, Yujing Wang, Peng Xing, Qinglong Wu
    International Journal of Systematic and Evolutionary Microbiology .2024;[Epub]     CrossRef
  • Saline soil improvement promotes the transformation of microbial salt tolerance mechanisms and microbial-plant-animal ecological interactions
    Keyu Yao, Guanghao Wang, Wen Zhang, Qiang Liu, Jian Hu, Mao Ye, Xin Jiang
    Journal of Environmental Management.2024; 372: 123360.     CrossRef
  • Optimal Irrigation and Fertilization Enhanced Tomato Yield and Water and Nitrogen Productivities by Increasing Rhizosphere Microbial Nitrogen Fixation
    Hongfei Niu, Tieliang Wang, Yongjiang Dai, Mingze Yao, Bo Li, Jiaqi Zheng, Lizhen Mao, Mingyu Zhao, Zhanyang Xu, Feng Zhang
    Agronomy.2024; 14(9): 2111.     CrossRef
  • Short-term effect of reclaimed wastewater quality gradient on soil microbiome during irrigation
    V. Moulia, N. Ait-Mouheb, G. Lesage, J. Hamelin, N. Wéry, V. Bru-Adan, L. Kechichian, M. Heran
    Science of The Total Environment.2023; 901: 166028.     CrossRef
  • Dyadobacter pollutisoli sp. nov., isolated from plastic waste landfill soil
    Kyeong Ryeol Kim, Jeong Min Kim, Jae Kyeong Lee, Dong Min Han, Lujiang Hao, Che Ok Jeon
    International Journal of Systematic and Evolutionary Microbiology .2023;[Epub]     CrossRef
  • Physiological and genomic analyses of cobalamin (vitamin B12)-auxotrophy of Lysobacter auxotrophicus sp. nov., a methionine-auxotrophic chitinolytic bacterium isolated from chitin-treated soil
    Akihiro Saito, Hideo Dohra, Moriyuki Hamada, Ryota Moriuchi, Yohei Kotsuchibashi, Koji Mori
    International Journal of Systematic and Evolutionary Microbiology .2023;[Epub]     CrossRef
  • Nitratireductor rhodophyticola sp. nov., isolated from marine red algae
    Kyung Hyun Kim, Sylvia Kristyanto, Hyung Min Kim, Kyeong Ryeol Kim, Che Ok Jeon
    International Journal of Systematic and Evolutionary Microbiology .2022;[Epub]     CrossRef
  • Description of Corynebacterium poyangense sp. nov., isolated from the feces of the greater white-fronted geese (Anser albifrons)
    Qian Liu, Guoying Fan, Kui Wu, Xiangning Bai, Xi Yang, Wentao Song, Shengen Chen, Yanwen Xiong, Haiying Chen
    Journal of Microbiology.2022; 60(7): 668.     CrossRef
  • Lysobacter ciconiae sp. nov., and Lysobacter avium sp. nov., isolated from the faeces of an Oriental stork
    So-Yeon Lee, Pil Soo Kim, Hojun Sung, Dong-Wook Hyun, Jin-Woo Bae
    Journal of Microbiology.2022; 60(5): 469.     CrossRef
  • Isolation and characterization of tick-borne Roseomonas haemaphysalidis sp. nov. and rodent-borne Roseomonas marmotae sp. nov.
    Wentao Zhu, Juan Zhou, Shan Lu, Jing Yang, Xin-He Lai, Dong Jin, Ji Pu, Yuyuan Huang, Liyun Liu, Zhenjun Li, Jianguo Xu
    Journal of Microbiology.2022; 60(2): 137.     CrossRef
  • Rhodococcus oxybenzonivorans sp. nov., a benzophenone-3-degrading bacterium, isolated from stream sediment
    Ju Hye Baek, Woonhee Baek, Sang Eun Jeong, Sung Chul Lee, Hyun Mi Jin, Che Ok Jeon
    International Journal of Systematic and Evolutionary Microbiology.2022;[Epub]     CrossRef
Full-repertoire comparison of the microscopic objects composing the human gut microbiome with sequenced and cultured communities
Edmond Kuete Yimagou , Jean-Pierre Baudoin , Rita Abou Abdallah , Fabrizio Di Pinto , Jacques Yaacoub Bou Khalil , Didier Raoult
J. Microbiol. 2020;58(5):377-386.   Published online April 11, 2020
DOI: https://doi.org/10.1007/s12275-020-9365-3
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AbstractAbstract
The study of the human gut microbiome is essential in microbiology and infectious diseases as specific alterations in the gut microbiome might be associated with various pathologies, such as chronic inflammatory disease, intestinal infection and colorectal cancer. To identify such dysregulations, several strategies are being used to create a repertoire of the microorganisms composing the human gut microbiome. In this study, we used the “microscomics” approach, which consists of creating an ultrastructural repertoire of all the cell-like objects composing stool samples from healthy donors using transmission electron microscopy (TEM). We used TEM to screen ultrathin sections of 8 resin-embedded stool samples. After exploring hundreds of micrographs, we managed to elaborate ultrastructural categories based on morphological criteria or features. This approach explained many inconsistencies observed with other techniques, such as metagenomics and culturomics. We highlighted the value of our cultureindependent approach by comparing our microscopic images to those of cultured bacteria and those reported in the literature. This study helped to detect “minimicrobes” Candidate Phyla Radiation (CPR) for the first time in human stool samples. This “microscomics” approach is non-exhaustive but complements already existing approaches and adds important data to the puzzle of the microbiota.

Citations

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  • Candidate Phyla Radiation, an Underappreciated Division of the Human Microbiome, and Its Impact on Health and Disease
    Sabrina Naud, Ahmad Ibrahim, Camille Valles, Mohamad Maatouk, Fadi Bittar, Maryam Tidjani Alou, Didier Raoult
    Clinical Microbiology Reviews.2022;[Epub]     CrossRef
  • Radiotherapy and the gut microbiome: facts and fiction
    Jing Liu, Chao Liu, Jinbo Yue
    Radiation Oncology.2021;[Epub]     CrossRef
  • Host–microbiota maladaptation in colorectal cancer
    Alina Janney, Fiona Powrie, Elizabeth H. Mann
    Nature.2020; 585(7826): 509.     CrossRef
Impact of small RNA RaoN on nitrosative-oxidative stress resistance and virulence of Salmonella enterica serovar Typhimurium
Sinyeon Kim , Yong Heon Lee
J. Microbiol. 2020;58(6):499-506.   Published online April 11, 2020
DOI: https://doi.org/10.1007/s12275-020-0027-2
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AbstractAbstract
RaoN is a Salmonella-specific small RNA that is encoded in the cspH-envE intergenic region on Salmonella pathogenicity island-11. We previously reported that RaoN is induced under conditions of acid and oxidative stress combined with nutrient limitation, contributing to the intramacrophage growth of Salmonella enterica serovar Typhimurium. However, the role of RaoN in nitrosative stress response and virulence has not yet been elucidated. Here we show that the raoN mutant strain has increased susceptibility to nitrosative stress by using a nitric oxide generating acidified nitrite. Extending previous research on the role of RaoN in oxidative stress resistance, we found that NADPH oxidase inhibition restores the growth of the raoN mutant in LPS-treated J774A.1 macrophages. Flow cytometry analysis further revealed that the inactivation of raoN leads to an increase in the intracellular level of reactive oxygen species (ROS) in Salmonella-infected macrophages, suggesting that RaoN is involved in the inhibition of NADPH oxidase-mediated ROS production by mechanisms not yet resolved. Moreover, we evaluated the effect of raoN mutation on the virulence in murine systemic infection and determined that the raoN mutant is less virulent than the wild-type strain following oral inoculation. In
conclusion
, small regulatory RNA RaoN controls nitrosativeoxidative stress resistance and is required for virulence of Salmonella in mice.

Citations

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  • The Salmonella enterica EnvE is an Outer Membrane Lipoprotein and Its Gene Expression Leads to Transcriptional Repression of the Virulence Gene msgA
    Sinyeon Kim, Yong Heon Lee
    Journal of Microbiology.2024; 62(11): 1013.     CrossRef
  • Functions of Small Non-Coding RNAs in Salmonella–Host Interactions
    Xia Meng, Mengping He, Pengpeng Xia, Jinqiu Wang, Heng Wang, Guoqiang Zhu
    Biology.2022; 11(9): 1283.     CrossRef
  • Detoxification Response of Pseudomonas fluorescens MFAF76a to Gaseous Pollutants NO2 and NO
    Thibault Chautrand, Ségolène Depayras, Djouhar Souak, Mathilde Bouteiller, Tatiana Kondakova, Magalie Barreau, Mohamed Amine Ben Mlouka, Julie Hardouin, Yoan Konto-Ghiorghi, Sylvie Chevalier, Annabelle Merieau, Nicole Orange, Cécile Duclairoir-Poc
    Microorganisms.2022; 10(8): 1576.     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
  • Gaseous NO2 induces various envelope alterations in Pseudomonas fluorescens MFAF76a
    Thibault Chautrand, Ségolène Depayras, Djouhar Souak, Tatiana Kondakova, Magalie Barreau, Takfarinas Kentache, Julie Hardouin, Ali Tahrioui, Olivier Thoumire, Yoan Konto-Ghiorghi, Corinne Barbey, Guy Ladam, Sylvie Chevalier, Hermann J. Heipieper, Nicole O
    Scientific Reports.2022;[Epub]     CrossRef
  • Current challenges facing one-step production of l-ascorbic acid
    Panpan Wang, Weizhu Zeng, Sha Xu, Guocheng Du, Jingwen Zhou, Jian Chen
    Biotechnology Advances.2018; 36(7): 1882.     CrossRef
Zur-regulated lipoprotein A contributes to the fitness of Acinetobacter baumannii
Eun Kyung Lee , Chul Hee Choi , Man Hwan Oh
J. Microbiol. 2020;58(1):67-77.   Published online January 2, 2020
DOI: https://doi.org/10.1007/s12275-020-9531-7
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AbstractAbstract
Acinetobacter baumannii is a notorious nosocomial pathogen that commonly infects severely ill patients. Zinc (Zn) is essential to survive and adapt to different environment and host niches in A. baumannii. Of the Zinc uptake regulator (Zur)-regulated genes in A. baumannii, the A1S_3412 gene encoding a Zur-regulated lipoprotein A (ZrlA) is critical for cell envelope integrity and overcoming antibiotic exposure. This study investigated whether ZrlA contributes to the fitness of A. baumannii in vitro and in vivo using the wildtype A. baumannii ATCC 17978, ΔzrlA mutant, and zrlAcomplemented strains. The ΔzrlA mutant showed reduced biofilm formation, surface motility, and adherence to and invasion of epithelial cells compared to the wild-type strain. In a mouse pneumonia model, the ΔzrlA mutant showed significantly lower bacterial numbers in the blood than the wildtype strain. These virulence traits were restored in the zrlAcomplemented strain. Under static conditions, the expression of csuCDE, which are involved in the chaperone-usher pili assembly system, was significantly lower in the ΔzrlA mutant than in the wild-type strain. Moreover, the expression of the bfmR/S genes, which regulate the CsuA/BABCDE system, was significantly lower in the ΔzrlA mutant under static conditions than in the wild-type strain. Our results indicate that the zrlA gene plays a role in the fitness of A. baumannii by regulating the BfmR/S two-component system and subsequently the CsuA/BABCDE chaperone-usher pili assembly system, suggesting it as a potential target for anti-virulence strategies against A. baumannii.

Citations

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  • Molecular Detection of Pap II, OmpA, and LuxR Genes Responsible for Biofilm Formation in Acinetobacter baumannii Isolated from Hospitalized Patients
    Estabraq Ali Maklef, Amal A. Kareem, Susan F. K. Al-Sudani
    Medical Journal of Babylon.2024; 21(Suppl 2): S258.     CrossRef
  • Pathogenicity and virulence of Acinetobacter baumannii : Factors contributing to the fitness in healthcare settings and the infected host
    Massimiliano Lucidi, Daniela Visaggio, Antonella Migliaccio, Giulia Capecchi, Paolo Visca, Francesco Imperi, Raffaele Zarrilli
    Virulence.2024;[Epub]     CrossRef
  • Characterization of the Zinc Uptake Repressor (Zur) from Acinetobacter baumannii
    Minyong Kim, My Tra Le, Lixin Fan, Courtney Campbell, Sambuddha Sen, Daiana A. Capdevila, Timothy L. Stemmler, David P. Giedroc
    Biochemistry.2024; 63(5): 660.     CrossRef
  • Acinetobacter Metabolism in Infection and Antimicrobial Resistance
    Xiaomei Ren, Lauren D. Palmer, Karen M. Ottemann
    Infection and Immunity.2023;[Epub]     CrossRef
  • A response regulator controls Acinetobacter baumannii virulence by acting as an indole receptor
    Binbin Cui, Quan Guo, Xia Li, Shihao Song, Mingfang Wang, Gerun Wang, Aixin Yan, Jianuan Zhou, Yinyue Deng, Marenda Wilson-Pham
    PNAS Nexus.2023;[Epub]     CrossRef
  • The role of quorum sensing, biofilm formation, and iron acquisition as key virulence mechanisms in Acinetobacter baumannii and the corresponding anti-virulence strategies
    Soffi Kei Kei Law, Hock Siew Tan
    Microbiological Research.2022; 260: 127032.     CrossRef
  • Carboxy-Terminal Processing Protease Controls Production of Outer Membrane Vesicles and Biofilm in Acinetobacter baumannii
    Rakesh Roy, Ren-In You, Chan-Hua Chang, Chiou-Ying Yang, Nien-Tsung Lin
    Microorganisms.2021; 9(6): 1336.     CrossRef
  • ppGpp signaling plays a critical role in virulence of Acinetobacter baumannii
    Kyeongmin Kim, Maidul Islam, Hye-won Jung, Daejin Lim, Kwangsoo Kim, Sung-Gwon Lee, Chungoo Park, Je Chul Lee, Minsang Shin
    Virulence.2021; 12(1): 2122.     CrossRef
  • COG0523 proteins: a functionally diverse family of transition metal-regulated G3E P-loop GTP hydrolases from bacteria to man
    Katherine A Edmonds, Matthew R Jordan, David P Giedroc
    Metallomics.2021;[Epub]     CrossRef
  • The role of Zur-regulated lipoprotein A in bacterial morphology, antimicrobial susceptibility, and production of outer membrane vesicles in Acinetobacter baumannii
    Nayeong Kim, Hyo Jeong Kim, Man Hwan Oh, Se Yeon Kim, Mi Hyun Kim, Joo Hee Son, Seung Il Kim, Minsang Shin, Yoo Chul Lee, Je Chul Lee
    BMC Microbiology.2021;[Epub]     CrossRef
  • Insights Into Mechanisms of Biofilm Formation in Acinetobacter baumannii and Implications for Uropathogenesis
    Jennifer M. Colquhoun, Philip N. Rather
    Frontiers in Cellular and Infection Microbiology.2020;[Epub]     CrossRef
Research Support, Non-U.S. Gov't
NOTE] Identification of Chaperones in Freeze Tolerance in Saccharomyces cerevisiae
Mahendran Chinnamara Naicker , I Seul Jo , Hana Im
J. Microbiol. 2012;50(5):882-887.   Published online November 4, 2012
DOI: https://doi.org/10.1007/s12275-012-2411-z
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AbstractAbstract
Exposure to low temperatures reduces protein folding rates and induces the cold denaturation of proteins. Considering the roles played by chaperones in facilitating protein folding and preventing protein aggregation, chaperones must exist that confer tolerance to cold stress. Here, yeast strains lacking individual chaperones were screened for reduced freezing tolerance. In total, 19 of 82 chaperone-deleted strains tested were more sensitive to freeze-thaw treatment than wild-type cells. The reintroduction of the respective chaperone genes into the deletion mutants recovered the freeze tolerance. The freeze sensitivity of the chaperone-knockout strains was also retained in the presence of 20% glycerol.
Retracted Publication
NOTE] Identification of the Vibrio vulnificus htpG Gene and Its Influence on Cold Shock Recovery
Slae Choi , Kyungku Jang , Seulah Choi , Hee-jee Yun , Dong-Hyun Kang
J. Microbiol. 2012;50(4):707-711.   Published online August 25, 2012
DOI: https://doi.org/10.1007/s12275-012-2294-z
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AbstractAbstract
An htpG gene encoding the heat shock protein HtpG was identified and cloned from Vibrio vulnificus. The deduced amino acid sequence of HtpG from V. vulnificus exhibited 71 and 85% identity to those reported from Escherichia coli and V. cholera, respectively. Functions of HtpG were assessed by the construction of an isogenic mutant whose htpG gene was deleted and by evaluating its phenotype changes during and after cold shock. The results demonstrated that recovery of the wild type from cold shock was significantly faster (p<0.05) than that of the htpG mutant, and indicated that the chaperone protein HtpG contributes to cold shock recovery, rather than cold shock tolerance, of V. vulnificus.
Journal Article
Transcriptional and Biochemical Characterization of Two Azotobacter vinelandii FKBP Family Members
Maria Dimou , Chrysoula Zografou , Anastasia Venieraki , Panagiotis Katinakis
J. Microbiol. 2011;49(4):635-640.   Published online September 2, 2011
DOI: https://doi.org/10.1007/s12275-011-0498-2
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AbstractAbstract
Peptidyl-prolyl cis/trans isomerases (PPIases, EC: 5.2.1.8), a class of enzymes that catalyse the rate-limiting step of the cis/trans isomerization in protein folding, are divided into three structurally unrelated families: cyclophilins, FK506-binding proteins (FKBPs), and parvulins. Two recombinant FKBPs from the soil nitrogenfixing bacterium Azotobacter vinelandii, designated as AvfkbX and AvfkbB, have been purified and their peptidyl-prolyl cis/trans isomerase activity against Suc-Ala-Xaa-Pro-Phe-pNA synthetic peptides characterised. The substrate specificity of both enzymes is typical for bacterial FKBPs, with Suc-Ala-Phe-Pro-Phe-pNA being the most rapidly catalysed substrate by AvfkbX and Suc-Ala-Leu-Pro-Phe-pNA by AvfkbB. Both FKBPs display chaperone activity as well in the citrate synthase thermal aggregation assay. Furthermore, using real-time RT-qPCR, we demonstrated that both genes were expressed during the exponential growth phase on glucose minimal medium, while their expression declined dramatically during the stationary growth phase as well as when the growth medium was supplied exogenously with ammonium.

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