Journal of Microbiology

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Cryptic prophages in a blaNDM-1-bearing plasmid increase bacterial survival against high NaCl concentration, high and low temperatures, and oxidative and immunological stressors: So Yeon Kim , Kwan Soo Ko; J. Microbiol. 2020;58(6):483-488. Published online March 28, 2020; DOI: https://doi.org/10.1007/s12275-020-9605-6

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In this study, we investigated the effect of cryptic prophage regions in a blaNDM-1-bearing plasmid, which was identified in a patient from South Korea, on the survival of bacteria against adverse environmental conditions. First, we conjugated the intact plasmid and plasmids with deleted cryptic prophages into Escherichia coli DH5α. The E. coli transconjugants carrying the plasmid with intact cryptic prophages showed increased survival during treatment with a high concentration of NaCl, high and low temperatures, an oxidative stressor (H2O2), and an immunological stressor (human serum). By contrast, the transconjugants carrying the plasmid with a single-cryptic prophage knockout did not show any change in survival rates. mRNA expression analyses revealed that the genes encoding sigma factor proteins were highly upregulated by the tested stressors and affected the expression of various proteins (antioxidant, cell osmosis-related, heat shock, cold shock, and universal stress proteins) associated with the specific defense against each stress. These findings indicate that a bacterial strain carrying a plasmid with intact carbapenemase gene and cryptic prophage regions exhibited an increased resistance against simulated environmental stresses, and cryptic prophages in the plasmid might contribute to this enhanced stress resistance. Our study indicated that the coselection of antibiotic resistance and resistance to other stresses may help bacteria to increase survival rates against adverse environments and disseminate.

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Journal Articles

Methyltransferase of a cell culture-adapted hepatitis E inhibits the MDA5 receptor signaling pathway: Jinjong Myoung , Jeong Yoon Lee , Kang Sang Min; J. Microbiol. 2019;57(12):1126-1131. Published online November 22, 2019; DOI: https://doi.org/10.1007/s12275-019-9478-8

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Abstract

Hepatitis E virus (HEV) is a causative agent of acute hepatitis and jaundice. The number of human infections is approximated to be over 20 million cases per year. The transmission is mainly via the fecal-oral route and contaminated water and food are considered to be a major source of infection. As a mouse model is not available, a recent development of a cell culture-adapted HEV strain (47832c) is considered as a very important tools for molecular analysis of HEV pathogenesis in cells. Previously, we demonstrated that HEV-encoded methyltransferase (MeT) encoded by the 47832c strain inhibits MDA5- and RIG-I-mediated activation of interferon β (IFN-β) promoter. Here, we report that MeT impairs the phosphorylation and activation of interferon regulatory factor 3 and the p65 subunit of NF-κB in a dose-dependent manner. In addition, the MeT encoded by the 47832c, but not that of HEV clinical or field isolates (SAR-55, Mex-14, KC-1, and ZJ-1), displays the inhibitory effect. A deeper understanding of MeTmediated suppression of IFN-β expression would provide basis of the cell culture adaptation of HEV.

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Viral Hepatitis: Host Immune Interaction, Pathogenesis and New Therapeutic Strategies
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Yannick Brüggemann, Mara Klöhn, Heiner Wedemeyer, Eike Steinmann
Nature Reviews Gastroenterology & Hepatology.2024; 21(10): 710. CrossRef
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Florencia Cancela, Ofelia Noceti, Juan Arbiza, Santiago Mirazo
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Host Innate Immunity Against Hepatitis Viruses and Viral Immune Evasion
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Frontiers in Microbiology.2021;[Epub] CrossRef
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Sameer-ul-Salam Mattoo, Jinjong Myoung
Journal of Microbiology and Biotechnology.2021; 31(12): 1601. CrossRef
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Sébastien Lhomme, Marion Migueres, Florence Abravanel, Olivier Marion, Nassim Kamar, Jacques Izopet
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Chikungunya Virus nsP2 Impairs MDA5/RIG-I-Mediated Induction of NF-κB Promoter Activation: A Potential Target for Virus-Specific Therapeutics
Sojung Bae, Jeong Yoon Lee, Jinjong Myoung
Journal of Microbiology and Biotechnology.2020; 30(12): 1801. CrossRef
Zika Virus-Encoded NS2A and NS4A Strongly Downregulate NF-κB Promoter Activity
Jeong Yoon Lee, Thi Thuy Ngan Nguyen, Jinjong Myoung
Journal of Microbiology and Biotechnology.2020; 30(11): 1651. CrossRef

Gentic overexpression increases production of hypocrellin A in Shiraia bambusicola S4201: Dan Li , Ning Zhao , Bing-Jing Guo , Xi Lin , Shuang-Lin Chen , Shu-Zhen Yan; J. Microbiol. 2019;57(2):154-162. Published online January 31, 2019; DOI: https://doi.org/10.1007/s12275-019-8259-8

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Abstract

Hypocrellin A (HA) is a perylenequinone (PQ) isolated from Shiraia bambusicola that shows antiviral and antitumor activities, but its application is limited by the low production from wild fruiting body. A gene overexpressing method was expected to augment the production rate of HA in S. bambusicola. However, the application of this molecular biology technology in S. bambusicola was impeded by a low genetic transformation efficiency and little genomic information. To enhance the plasmid transformant ratio, the Polyethylene Glycol-mediated transformation system was established and optimized. The following green fluorescent protein (GFP) analysis showed that the gene fusion expression system we constructed with a GAPDH promoter Pgpd1 and a rapid 2A peptide was successfully expressed in the S. bambusicola S4201 strain. We successfully obtained the HA high-producing strains by overexpressing O-methyltransferase/FAD-dependent monooxygenase gene (mono) and the hydroxylase gene (hyd), which were the essential genes involved in our putative HA biosynthetic pathway. The overexpression of these two genes increased the production of HA by about 200% and 100%, respectively. In general, this study will provide a basis to identify the genes involved in the hypocrellin A biosynthesis. This improved transformation method can also be used in genetic transformation studies of other fungi.

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Research Support, Non-U.S. Gov't

Identification of a Methyltransferase Encoded by Gene ste16 and Its Function in Ebosin Biosynthesis of Streptomyces sp. 139: Hong-Guan Xie , Yong-Gang Bao , Li-ping Bai , Jun-Jie Shan , Rong Jiang , Yang Zhang , Lian-Hong Guo , Ren Zhang , Yuan Li; J. Microbiol. 2009;47(2):193-200. Published online May 2, 2009; DOI: https://doi.org/10.1007/s12275-008-0195-y

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Abstract: Streptomyces sp. 139 generates a novel exopolysaccharide (EPS) designated as Ebosin, which exerts an antagonistic effect on IL-1R in vitro and anti-rheumatic arthritis activity in vivo. A ste gene cluster for Ebosin biosynthesis consisting of 27 ORFs was previously identified in our laboratory. In this paper, ste16 was expressed in Escherichia coli BL21 and the recombinant protein was purified, which has the ability to catalyze the transfer of the methyl group from S-adenosylmethionine (AdoMet) to dTDP-4-keto-6-deoxy-D-glucos, which was thus identified as a methyltransferase. In order to determine the function of ste16 in Ebosin biosynthesis, the gene was disrupted with a double crossover via homologous recombination. The monosaccharide composition of EPS-m generated by the mutant strain Streptomyces sp. 139 (ste16-) was found to differ from that of Ebosin. The IL-1R antagonist activity of EPS-m was markedly lower than that of Ebosin. These experimental results have shown that the ste16 gene codes for a methyltransferase which is involved in Ebosin biosynthesis.

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