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Volume 60(9); September 2022
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[PROTOCOL]Analyzing viral epitranscriptomes using nanopore direct RNA sequencing
Ari Hong , Dongwan Kim , V. Narry Kim , Hyeshik Chang
J. Microbiol. 2022;60(9):867-876.   Published online August 24, 2022
DOI: https://doi.org/10.1007/s12275-022-2324-4
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AbstractAbstract
RNA modifications are a common occurrence across all domains of life. Several chemical modifications, including N6- methyladenosine, have also been found in viral transcripts and viral RNA genomes. Some of the modifications increase the viral replication efficiency while also helping the virus to evade the host immune system. Nonetheless, there are numerous examples in which the host's RNA modification enzymes function as antiviral factors. Although established methods like MeRIP-seq and miCLIP can provide a transcriptome- wide overview of how viral RNA is modified, it is difficult to distinguish between the complex overlapping viral transcript isoforms using the short read-based techniques. Nanopore direct RNA sequencing (DRS) provides both long reads and direct signal readings, which may carry information about the modifications. Here, we describe a refined protocol for analyzing the RNA modifications in viral transcriptomes using nanopore technology.

Citations

Citations to this article as recorded by  
  • Utilization of nanopore direct RNA sequencing to analyze viral RNA modifications
    Lu Tan, Zhihao Guo, Xiaoming Wang, Dal Young Kim, Runsheng Li, Pedro H. Oliveira
    mSystems.2024;[Epub]     CrossRef
  • Current progress in strategies to profile transcriptomic m6A modifications
    Yuening Yang, Yanming Lu, Yan Wang, Xianghui Wen, Changhai Qi, Weilan Piao, Hua Jin
    Frontiers in Cell and Developmental Biology.2024;[Epub]     CrossRef
  • Improved sub-genomic RNA prediction with the ARTIC protocol
    Thomas Baudeau, Kristoffer Sahlin
    Nucleic Acids Research.2024; 52(17): e82.     CrossRef
  • Non-Targeted RNA Sequencing: Towards the Development of Universal Clinical Diagnosis Methods for Human and Veterinary Infectious Diseases
    Stephen Spatz, Claudio L. Afonso
    Veterinary Sciences.2024; 11(6): 239.     CrossRef
  • Quantitative profiling N1-methyladenosine (m1A) RNA methylation from Oxford nanopore direct RNA sequencing data
    Shenglun Chen, Jia Meng, Yuxin Zhang
    Methods.2024; 228: 30.     CrossRef
  • Multicellular, IVT-derived, unmodified human transcriptome for nanopore-direct RNA analysis
    Caroline A. McCormick, Stuart Akeson, Sepideh Tavakoli, Dylan Bloch, Isabel N. Klink, Miten Jain, Sara H. Rouhanifard
    Gigabyte.2024; 2024: 1.     CrossRef
  • Dissecting the effects of METTL3 on alternative splicing in prostate cancer
    Lin Wang, Ling Shi, Yonghao Liang, Judy Kin-Wing Ng, Chan Hoi Yin, Lingyi Wang, Jinpao Hou, Yiwei Wang, Cathy Sin-Hang Fung, Peter Ka-Fung Chiu, Chi-Fai Ng, Stephen Kwok-Wing Tsui
    Frontiers in Oncology.2023;[Epub]     CrossRef
Characterization of antibiotic-resistant, coagulase-negative staphylococci from fresh produce and description of Staphylococcus shinii sp. nov. isolated from chives
Gyu-Sung Cho , Bo Li , Erik Brinks , Charles , M.A.P. Franz
J. Microbiol. 2022;60(9):877-889.   Published online June 22, 2022
DOI: https://doi.org/10.1007/s12275-022-2100-5
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AbstractAbstract
Coagulase-negative Staphylococcus (CoNS) species may possess antibiotic resistance genes and have been associated with nosocomial infections. In this study, 91 CoNS with decreased susceptibility to oxacillin were isolated from fresh produce using oxacillin containing agar plates. Their antibiotic resistances were determined phenotypically and all isolates were identified by rep-PCR, 16S rRNA and rpoB gene sequencing. Furthermore, the genomes of representative strains were sequenced in order to confirm species identification by phylogenomics. The majority (64 of 91) of the CoNS strains could be identified as Mammaliicoccus (M.) fleurettii, while 13 were identified as M. sciuri, 8 as M. vitulinus, 2 as Staphylococcus (S.) epidermidis and single strains each as S. warneri, S. xylosus, Staphylococcus spp. and S. casei. Most of the strains were generally susceptible to clinically-relevant antibiotics, but only few (< 7%) strains possessed multiple resistances. Both oxacillin and cefoxitin resistant isolates were considered to be presumptive methicillin-resistant CoNS. From whole genome sequencing data of 6 representative strains, the mecA gene, accessory genes and the SCC loci were compared, which revealed high variability between some of the strains. The major fatty acids of K22-5MT strain included anteiso-C15:0, iso-C15:0, iso-C17:0, anteiso-C17:0, C18:0, and C20:0. Average nucleotide identity and digital DNA-DNA hybridization values indicated that Staphylococcus strain K22-5MT was below the species delineation cutoff values for ANI (less than 91%) and DDH (less than 44.4%), with the most closely related species being the S. pseudoxylosus S04009T type strain. Thus, strain K22- 5MT (=DSM 112532T, =LMG 32324T) represents a novel species, for which the name Staphylococcus shinii sp. nov. is proposed.

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  • Microbial agents for the control of ticks Rhipicephalus microplus
    Edgar Castro-Saines, Rodolfo Lagunes-Quintanilla, Rubén Hernández-Ortiz
    Parasitology Research.2024;[Epub]     CrossRef
  • Scandinavium lactucae sp. nov. Isolated from Healthy Lettuce in South Korea
    Jiwon Park, Sieun Park, Kwang-Kyo Oh, Charles M. A. P. Franz, Gyu-Sung Cho
    Current Microbiology.2024;[Epub]     CrossRef
  • Efficacy of Entomopathogenic Staphylococcus Bacteria as a Biocontrol Agent against Rhipicephalus microplus Ticks: Assessing Reproductive Inhibition and Mortality Rates
    Raquel Cossio-Bayugar, Cesar A. Arreguin-Perez, Hugo Aguilar-Diaz, Estefan Miranda-Miranda
    Microorganisms.2024; 12(3): 551.     CrossRef
  • Genomic exploration of the fermented meat isolate Staphylococcus shinii IMDO-S216 with a focus on competitiveness-enhancing secondary metabolites
    Ana Sosa-Fajardo, Cristian Díaz-Muñoz, David Van der Veken, Inés Pradal, Marko Verce, Stefan Weckx, Frédéric Leroy
    BMC Genomics.2024;[Epub]     CrossRef
  • Staphylococcus hsinchuensis sp. nov., Isolated from Soymilk
    Yu-Ting Wang, Yu-Chun Lin, Yi-Huei Hsieh, Yu-Tzu Lin, Moriyuki Hamada, Chih-Chieh Chen, Jong-Shian Liou, Ai-Yun Lee, Wei-Ling Zhang, Yung-Tsung Chen, Chien-Hsun Huang
    Pathogens.2024; 13(4): 343.     CrossRef
  • Validation List no. 209. Valid publication of new names and new combinations effectively published outside the IJSEM
    Aharon Oren, Markus Göker
    International Journal of Systematic and Evolutionary Microbiology .2023;[Epub]     CrossRef
Description of Flavobacterium cyclinae sp. nov. and Flavobacterium channae sp. nov., isolated from the intestines of Cyclina sinensis (Corb shell) and Channa argus (Northern snakehead)
Seomin Kang , Jae-Yun Lee , Jeong Eun Han , Yun-Seok Jeong , Do-Hun Gim , Jin-Woo Bae
J. Microbiol. 2022;60(9):890-898.   Published online June 22, 2022
DOI: https://doi.org/10.1007/s12275-022-2075-2
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AbstractAbstract
Two novel bacterial strains, KSM-R2A25T and KSM-R2A30T, were isolated from intestines of Cyclina sinensis (corb shell) and Channa argus (northern snakehead), respectively. Both specimens were collected in Korea. The strains were Gramstain- negative, non-motile, and strictly aerobic. According to phylogenetic analyses based on 16S rRNA gene sequences, strains belonged to the genus Flavobacterium within the family Flavobacteriaceae. 16S rRNA gene sequences of strains KSMR2A25T and KSM-R2A30T were closely related to Flavobacterium cucumis DSM 18830T and Flavobacterium aquaticum JC164T with sequence similarities of 97.77% and 98.54%, respectively. Further genomic analyses including reconstruction of the UBCG tree and overall genome-related indices suggested them as novel species of the genus Flavobacterium. Both strains contained menaquinone with six isoprene units (MK-6) as a major isoprenoid quinone and iso-C15:1 G, iso- C15:0, and iso-C16:0 as major cellular fatty acids. The major polar lipid in both strains was phosphatidylethanolamine. The genomic G + C contents of strains KSM-R2A25T and KSMR2A30T were 31.7 and 31.9%, respectively. Based on the polyphasic taxonomic study presented here, strains KSM-R2A25T and KSM-R2A30T represent novel species of the genus Flavobacterium, for which the names Flavobacterium cyclinae sp. nov and Flavobacterium channae sp. nov are proposed. The type strains of F. cyclinae sp. nov and F. channae sp. nov are KSM-R2A25T (= KCTC 82978T = JCM 34997T) and KSMR2A30T (= KCTC 82979T = JCM 34998T), respectively.

Citations

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  • Flavobacterium rivulicola sp. nov., Isolated from a Freshwater Stream
    Sumin Kim, Miri S. Park, Ilnam Kang, Jang-Cheon Cho
    Current Microbiology.2024;[Epub]     CrossRef
  • Comprehensive genome analysis of five novel flavobacteria: Flavobacterium piscisymbiosum sp. nov., Flavobacterium pisciphilum sp. nov., Flavobacterium flavipigmentatum sp. nov., Flavobacterium lipolyticum sp. nov. and Flavobacterium cupriresistens sp. nov
    Izzet Burcin Saticioglu, Hilal Ay, Soner Altun, Nihed Ajmi, Enes Said Gunduz, Huban Gocmen, Muhammed Duman
    Systematic and Applied Microbiology.2024; 47(4): 126518.     CrossRef
  • The first report of submandibular lymph node infection with Flavobacterium ceti in a horse and its complete genome sequence
    Seung-Ho Ryu, Beomsoon Jang, Byung-Sun Kim, Kun Taek Park
    Microbial Pathogenesis.2024; 197: 107096.     CrossRef
  • Rhodobacteraceae are Prevalent and Ecologically Crucial Bacterial Members in Marine Biofloc Aquaculture
    Meora Rajeev, Jang-Cheon Cho
    Journal of Microbiology.2024; 62(11): 985.     CrossRef
  • Congregibacter variabilis sp. nov. and Congregibacter brevis sp. nov. Within the OM60/NOR5 Clade, Isolated from Seawater, and Emended Description of the Genus Congregibacter
    Hyeonsu Tak, Miri S. Park, Hyerim Cho, Yeonjung Lim, Jang-Cheon Cho
    Journal of Microbiology.2024; 62(9): 739.     CrossRef
  • An update on novel taxa and revised taxonomic status of bacteria isolated from aquatic host species described in 2022–2023
    Claire R. Burbick, Sara D. Lawhon, Brittany Bukouras, Giovanna Lazzerini, Erik Munson, Romney M. Humphries
    Journal of Clinical Microbiology.2024;[Epub]     CrossRef
  • Transcriptomic insights into the immune response of the intestine to Aeromonas veronii infection in northern snakehead (Channa argus)
    Peng Sun, Di Zhang, Na Li, Xiao-Fei Li, Yi-Han Ma, Hui Li, Ye Tian, Tao Wang, Shahrood Ahmed Siddiquid, Wu-wen Sun, Lei Zhang, Xiao-Feng Shan, Chun-Feng Wang, Ai-Dong Qian, Dong-Xing Zhang
    Ecotoxicology and Environmental Safety.2023; 255: 114825.     CrossRef
  • Ten Novel Species Belonging to the Genus Flavobacterium, Isolated from Freshwater Environments: F. praedii sp. nov., F. marginilacus sp. nov., F. aestivum sp. nov., F. flavigenum sp. nov., F. luteolum sp. nov., F. gelatinilyticum sp. nov., F. aquiphilum s
    Hyunyoung Jo, Miri S. Park, Yeonjung Lim, Ilnam Kang, Jang-Cheon Cho
    Journal of Microbiology.2023; 61(5): 495.     CrossRef
  • Validation List no. 209. Valid publication of new names and new combinations effectively published outside the IJSEM
    Aharon Oren, Markus Göker
    International Journal of Systematic and Evolutionary Microbiology .2023;[Epub]     CrossRef
Sala cibi gen. nov., sp. nov., an extremely halophilic archaeon isolated from solar salt
Hye Seon Song , Juseok Kim , Yeon Bee Kim , Se Hee Lee , Tae Woong Whon , Seong Woon Roh
J. Microbiol. 2022;60(9):899-904.   Published online July 14, 2022
DOI: https://doi.org/10.1007/s12275-022-2137-5
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AbstractAbstract
Two novel halophilic archaeal strains, CBA1133T and CBA- 1134, were isolated from solar salt in South Korea. The 16S rRNA gene sequences of the isolates were identical to each other and were closely related to the genera Natronomonas (92.3–93.5%), Salinirubellus (92.2%), Halomarina (91.3– 92.0%), and Haloglomus (91.4%). The isolated strains were coccoid, Gram-stain-negative, aerobic, oxidase-positive, and catalase-negative. Growth occurred under temperatures of 25–50°C (optimum, 45°C), NaCl levels of 10–30% (optimum, 15%), pH levels of 6.0–8.5 (optimum, 7.0), and MgCl2 concentrations of 0–500 mM (optimum, 100 mM). Digital DNADNA hybridization values between the strains and related genera ranged from 18.3% to 22.7%. The major polar lipids of the strains were phosphatidyl glycerol, phosphatidyl glycerol phosphate methyl ester, and phosphatidyl glycerol sulfate. Genomic, phenotypic, physiological, and biochemical analyses of the isolates revealed that they represent a novel genus and species in the family Halobacteriaceae. The type strain is CBA1133T (= KACC 22148T = JCM 34265T), for which the name Sala cibi gen. nov., sp. nov. is proposed.

Citations

Citations to this article as recorded by  
  • Dominance and diversity of archaea in food-grade salts: insights for histamine degradation in salt-fermented foods
    Jing Hou, Ya-Ling Mao, Na Li, Xiao-Yan Yang, Chidiebele Nwankwo, Heng-Lin Cui
    International Journal of Food Science and Technology.2024; 59(12): 9490.     CrossRef
  • Congregibacter variabilis sp. nov. and Congregibacter brevis sp. nov. Within the OM60/NOR5 Clade, Isolated from Seawater, and Emended Description of the Genus Congregibacter
    Hyeonsu Tak, Miri S. Park, Hyerim Cho, Yeonjung Lim, Jang-Cheon Cho
    Journal of Microbiology.2024; 62(9): 739.     CrossRef
  • Genome-based classification of genera Halosegnis and Salella, and description of four novel halophilic archaea isolated from a tidal flat
    Yao Hu, Xue Ma, Shun Tan, Xin-Xin Li, Mu Cheng, Jing Hou, Heng-Lin Cui
    Antonie van Leeuwenhoek.2024;[Epub]     CrossRef
  • Assessing the impact of heavy metals on bacterial diversity in coastal regions of Southeastern India
    Chandra Veluchamy, Avinash Sharma, Kalaivani Thiagarajan
    Environmental Monitoring and Assessment.2024;[Epub]     CrossRef
  • Proposal of Eoetvoesiella gen. nov., Paludihabitans gen. nov., Rivihabitans gen. nov. and Salella gen. nov. as replacement names for the illegitimate prokaryotic generic names Eoetvoesia, Paludicola, Rivicola and Sala, respectively
    Umakant Bhoopati Deshmukh, Aharon Oren
    International Journal of Systematic and Evolutionary Microbiology .2023;[Epub]     CrossRef
  • Validation List no. 209. Valid publication of new names and new combinations effectively published outside the IJSEM
    Aharon Oren, Markus Göker
    International Journal of Systematic and Evolutionary Microbiology .2023;[Epub]     CrossRef
  • Halorarius litoreus gen. nov., sp. nov., Halorarius halobius sp. nov., Haloglomus halophilum sp. nov., Haloglomus salinum sp. nov., and Natronomonas marina sp. nov., extremely halophilic archaea isolated from tidal flat and marine solar salt
    Ya-Ping Sun, Bei-Bei Wang, Zhang-Ping Wu, Xi-Wen Zheng, Jing Hou, Heng-Lin Cui
    Frontiers in Marine Science.2023;[Epub]     CrossRef
  • Naming new taxa of prokaryotes in the 21st century
    Aharon Oren
    Canadian Journal of Microbiology.2023; 69(4): 151.     CrossRef
Characterization of Marinilongibacter aquaticus gen. nov., sp. nov., a unique marine bacterium harboring four CRISPR-Cas systems in the phylum Bacteroidota
Dao-Feng Zhang , Yu-Fang Yao , Hua-Peng Xue , Zi-Yue Fu , Xiao-Mei Zhang , Zongze Shao
J. Microbiol. 2022;60(9):905-915.   Published online August 1, 2022
DOI: https://doi.org/10.1007/s12275-022-2102-3
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AbstractAbstract
A novel bacterium, designated YYF0007T, was isolated from an agar-degrading co-culture. The strain was found harboring four CRISPR-Cas systems of two classes in the chromosome and subsequently subjected to a study on polyphasic taxonomy. Pairwise analyses of the 16S rRNA gene sequences indicated that strain YYF0007T had highest 16S rRNA gene sequence similarity (92.2%) to Jiulongibacter sediminis JN- 14-9T. The phylogenomic trees based on the 16S rRNA gene and 269 single-copy orthologous gene clusters (OCs) indicated that strain YYF0007T should be recognized as a novel genus of the family Spirosomaceae. The cells were Gramstain- negative, nonmotile, strictly aerobic, and straight long rods with no flagellum. Optimum growth occurred at 28°C and pH 7.0 with the presence of NaCl concentration 1.0–3.0% (w/v). The strain showed oxidase and catalase activities. The major fatty acids were C16:1ω5c, iso-C15:0 and summed feature 3 (C16:1 ω7c and/or C16:1 ω6c). The predominant isoprenoid quinone was MK-7. The complete genome size was 4.64 Mb with a DNA G + C content of 44.4%. Further typing of CRISPR-Cas systems in the family Spirosomaceae and the phylum Bacteroidota indicated that it was remarkable for strain YYF0007T featured by such a set of CRISPR-Cas systems. This trait highlights the applications of strain YYF- 0007T in studies on the evolutionary dynamics and bacterial autoimmunity of CRISPR-Cas system as a potential model. The name Marinilongibacter aquaticus gen. nov., sp. nov. is proposed, and the type strain is YYF0007T (= MCCC 1K06017T = GDMCC 1.2428T = JCM 34683T).

Citations

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  • Clustered Regularly Interspaced Short Palindromic Repeat/CRISPR-Associated Protein and Its Utility All at Sea: Status, Challenges, and Prospects
    Jiashun Li, Shuaishuai Wu, Kaidian Zhang, Xueqiong Sun, Wenwen Lin, Cong Wang, Senjie Lin
    Microorganisms.2024; 12(1): 118.     CrossRef
  • Unraveling the mechanisms behind sodium persulphate-induced changes in petroleum-contaminated aquifers’ biogeochemical parameters and microbial communities
    Yuqi Qi, Jun Zeng, Junshi Tao, Rentao Liu, Renchuan Fu, Chao Yan, Xiao Liu, Na Liu, Yanru Hao
    Chemosphere.2024; 351: 141174.     CrossRef
  • Arcicella gelida sp. nov. and Arcicella lustrica sp. nov., isolated from streams in China and re-examining the taxonomic status of all the genera within the families Spirosomataceae and Cytophagaceae
    Huibin Lu, Li Chen, Linpei Huang
    International Journal of Systematic and Evolutionary Microbiology .2024;[Epub]     CrossRef
  • Thalassospira aquimaris sp. nov. and Winogradskyella marincola sp. nov. two marine bacteria isolated from an agar-degrading co-culture
    Zi-Yue Fu, Dao-Feng Zhang, Meng-Han Huang, Hong-Chuan Wang, Xiao-Ye Chen, Yu-Fang Yao, Yang Yuan, Wen-Jun Li
    Antonie van Leeuwenhoek.2024;[Epub]     CrossRef
  • Validation List no. 209. Valid publication of new names and new combinations effectively published outside the IJSEM
    Aharon Oren, Markus Göker
    International Journal of Systematic and Evolutionary Microbiology .2023;[Epub]     CrossRef
  • Telluribacter roseus sp. nov., Isolated from the Kumtag Desert Soil
    Chu-Ying Feng, Jia-Rui Han, Chun-Yan Lu, Li Gu, Shuai Li, Wen-Hui Lian, Lei Dong, Wen-Jun Li
    Current Microbiology.2023;[Epub]     CrossRef
Complete gammaproteobacterial endosymbiont genome assembly from a seep tubeworm Lamellibrachia satsuma
Ajit Kumar Patra , Yong min Kwon , Youngik Yang
J. Microbiol. 2022;60(9):916-927.   Published online August 1, 2022
DOI: https://doi.org/10.1007/s12275-022-2057-4
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AbstractAbstract
Siboglinid tubeworms thrive in hydrothermal vent and seep habitats via a symbiotic relationship with chemosynthetic bacteria. Difficulties in culturing tubeworms and their symbionts in a laboratory setting have hindered the study of host-microbe interactions. Therefore, released symbiont genomes are fragmented, thereby limiting the data available on the genome that affect subsequent analyses. Here, we present a complete genome of gammaproteobacterial endosymbiont from the tubeworm Lamellibrachia satsuma collected from a seep in Kagoshima Bay, assembled using a hybrid approach that combines sequences generated from the Illumina and Oxford Nanopore platforms. The genome consists of a single circular chromosome with an assembly size of 4,323,754 bp and a GC content of 53.9% with 3,624 protein-coding genes. The genome is of high quality and contains no assembly gaps, while the completeness and contamination are 99.33% and 2.73%, respectively. Comparative genome analysis revealed a total of 1,724 gene clusters shared in the vent and seep tubeworm symbionts, while 294 genes were found exclusively in L. satsuma symbionts such as transposons, genes for defense mechanisms, and inorganic ion transportations. The addition of this complete endosymbiont genome assembly would be valuable for comparative studies particularly with tubeworm symbiont genomes as well as with other chemosynthetic microbial communities.

Citations

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  • Genomic and transcriptomic analyses illuminate the molecular basis of the unique lifestyle of a tubeworm, Lamellibrachia satsuma
    Taiga Uchida, Yuki Yoshioka, Yu Yoshida, Manabu Fujie, Ayuta Yamaki, Akira Sasaki, Koji Inoue, Chuya Shinzato
    DNA Research.2023;[Epub]     CrossRef
Characterization of transcriptional activities at a divergent promoter of the type VI secretion system in enterohemorrhagic Escherichia coli O157:H7
Se Kye Kim , Jun Bong Lee , Jang Won Yoon
J. Microbiol. 2022;60(9):928-934.   Published online August 19, 2022
DOI: https://doi.org/10.1007/s12275-022-2109-9
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AbstractAbstract
The type VI secretion system (T6SS) is a novel secretion system found in many Gram-negative bacteria that plays a role in bacterial competition, virulence, and host immune evasion. The enterohemorrhagic Escherichia coli (EHEC) O157:H7 strain EDL933 has a single functional T6SS gene cluster. In this study, we attempted to characterize the transcriptional pattern of the T6SS effector gene Z0264 in EDL933. Transcriptional analyses showed that Z0264 and other T6SS genes were transcribed in vitro in a growth-phase-dependent manner, but Z0264 was not secreted in the rich medium. Using adapter- and radioactivity-free transcription start site analysis, we identified an overlapping divergent promoter between Z0264 and Z0265. A β-galactosidase assay with truncated promoter regions showed that the divergent promoter is functional. In addition, we demonstrated the role of H-NS as a repressor in the transcription of Z0264. Notably, the cDNA PCR assay showed that the mRNA transcript from the Z0264 promoter did not include the entire main T6SS cluster, suggesting segmented gene expression by multiple promoters in the T6SS cluster. In conclusion, we identified a divergent promoter for Z0264 located in the T6SS cluster of EDL933 and characterized its in vitro transcriptional activity during growth. Our findings provide insights and a preliminary understanding of the regulatory mechanisms underlying T6SS transcription.
Activation of the SigE-SigB signaling pathway by inhibition of the respiratory electron transport chain and its effect on rifampicin resistance in Mycobacterium smegmatis
Yuna Oh , Hye-In Lee , Ji-A Jeong , Seonghan Kim , Jeong-Il Oh
J. Microbiol. 2022;60(9):935-947.   Published online August 1, 2022
DOI: https://doi.org/10.1007/s12275-022-2202-0
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AbstractAbstract
Using a mutant of Mycobacterium smegmatis lacking the major aa3 cytochrome c oxidase of the electron transport chain (Δaa3), we demonstrated that inhibition of the respiratory electron transport chain led to an increase in antibiotic resistance of M. smegmatis to isoniazid, rifampicin, ethambutol, and tetracycline. The alternative sigma factors SigB and SigE were shown to be involved in an increase in rifampicin resistance of M. smegmatis induced under respiration-inhibitory conditions. As in Mycobacterium tuberculosis, SigE and SigB form a hierarchical regulatory pathway in M. smegmatis through SigE-dependent transcription of sigB. Expression of sigB and sigE was demonstrated to increase in the Δaa3 mutant, leading to upregulation of the SigB-dependent genes in the mutant. The phoU2 (MSMEG_1605) gene implicated in a phosphatesignaling pathway and the MSMEG_1097 gene encoding a putative glycosyltransferase were identified to be involved in the SigB-dependent enhancement of rifampicin resistance observed for the Δaa3 mutant of M. smegmatis. The significance of this study is that the direct link between the functionality of the respiratory electron transport chain and antibiotic resistance in mycobacteria was demonstrated for the first time using an electron transport chain mutant rather than inhibitors of electron transport chain.

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  • Rel-dependent decrease in the expression of ribosomal protein genes by inhibition of the respiratory electron transport chain in Mycobacterium smegmatis
    Na-Kyeong Kim, Jong-Eun Baek, Ye-Jin Lee, Yuna Oh, Jeong-Il Oh
    Frontiers in Microbiology.2024;[Epub]     CrossRef
  • MoaB2, a newly identified transcription factor, binds to σ A in Mycobacterium smegmatis
    Barbora Brezovská, Subhash Narasimhan, Michaela Šiková, Hana Šanderová, Tomáš Kovaľ, Nabajyoti Borah, Mahmoud Shoman, Debora Pospíšilová, Viola Vaňková Hausnerová, Dávid Tužinčin, Martin Černý, Jan Komárek, Martina Janoušková, Milada Kambová, Petr Halada,
    Journal of Bacteriology.2024;[Epub]     CrossRef
  • Enhanced hypoxanthine utilization for cAMP salvage synthesis efficiently by Arthrobacter sp. CCTCC 2013431 via xanthine oxidase inhibition
    Baofeng Chen, Hai Tan, Chang Li, Linbo Li, Zhonghua Zhang, Zhigang Li
    Biotechnology Letters.2024; 46(6): 1095.     CrossRef
  • Mycobacterial Regulatory Systems Involved in the Regulation of Gene Expression Under Respiration-Inhibitory Conditions
    Yuna Oh, Ha-Na Lee, Eon-Min Ko, Ji-A Jeong, Sae Woong Park, Jeong-Il Oh
    Journal of Microbiology.2023; 61(3): 297.     CrossRef
Deletion of lacD gene affected stress tolerance and virulence of Streptococcus suis serotype 2
Xiaowu Jiang , Lexin Zhu , Dongbo Zhan
J. Microbiol. 2022;60(9):948-959.   Published online August 19, 2022
DOI: https://doi.org/10.1007/s12275-022-2146-4
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AbstractAbstract
Streptococcus suis type 2 (S. suis type 2, SS2), an infectious pathogen which is zoonotic and can induce severely public health concern. Our previous research identified a newly differential secreted effector of tagatose-bisphosphate aldolase (LacD) mediated by VirD4 factor within the putative type IV secretion system of SS2, whereas the functional basis and roles in virulence of LacD remain elusive. Here in this study, the LacD was found enzymatic and can be activated to express under oxidative stress. Gene mutant and its complemental strain (ΔlacD and cΔlacD) were constructed to analyze the phenotypes, virulence and transcriptomic profiles as compared with the parental strain. The lacD gene deletion showed no effect on growth capability and cells morphology of SS2. However, reduced tolerance to oxidative and heat stress conditions, increased antimicrobial susceptibility to ciprofloxacin and kanamycin were found in ΔlacD strain. Further, the LacD deficiency led to weakened invasion and attenuated virulence since an easier phagocytosed and more prone to be cleared of SS2 in macrophages were shown in ΔlacD mutant. Distinctive transcriptional profiling in ΔlacD strain and typical downregulated genes with significant mRNA changes including alcohol dehydrogenase, GTPase, integrative and conjugative elements, and iron ABC transporters which were mainly involved in cell division, stress response, antimicrobial susceptibility and virulence regulation, were examined and confirmed by RNA sequencing and real time qPCR. In summary, the
results
demonstrated for the first time that LacD was a pluripotent protein mediated the metabolic, stress and virulent effect of SS2.

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Enhancement of the solubility of recombinant proteins by fusion with a short-disordered peptide
Jun Ren , Suhee Hwang , Junhao Shen , Hyeongwoo Kim , Hyunjoo Kim , Jieun Kim , Soyoung Ahn , Min-gyun Kim , Seung Ho Lee , Dokyun Na
J. Microbiol. 2022;60(9):960-967.   Published online July 14, 2022
DOI: https://doi.org/10.1007/s12275-022-2122-z
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AbstractAbstract
In protein biotechnology, large soluble fusion partners are widely utilized for increased yield and solubility of recombinant proteins. However, the production of additional large fusion partners poses an additional burden to the host, leading to a decreased protein yield. In this study, we identified two highly disordered short peptides that were able to increase the solubility of an artificially engineered aggregationprone protein, GFP-GFIL4, from 0.6% to 61% (D3-DP00592) and 46% (D4-DP01038) selected from DisProt database. For further confirmation, the peptides were applied to two insoluble E. coli proteins (YagA and YdiU). The peptides also enhanced solubility from 52% to 90% (YagA) and from 27% to 93% (YdiU). Their ability to solubilize recombinant proteins was comparable with strong solubilizing tags, maltosebinding protein (40 kDa) and TrxA (12 kDa), but much smaller (< 7 kDa) in size. For practical application, the two peptides were fused with a restriction enzyme, I-SceI, and they increased I-SceI solubility from 24% up to 75%. The highly disordered peptides did not affect the activity of I-SceI while I-SceI fused with MBP or TrxA displayed no restriction activity. Despite the small size, the highly disordered peptides were able to solubilize recombinant proteins as efficiently as conventional fusion tags and did not interfere with the function of recombinant proteins. Consequently, the identified two highly disordered peptides would have practical utility in protein biotechnology and industry.

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