Journal of Microbiology

Hyung-Lyun Kang

6 Articles

Novel nuclear targeting coiled-coil protein of Helicobacter pylori showing Ca2+-independent, Mg2+-dependent DNase I activity: Young Chul Kwon , Sinil Kim , Yong Seok Lee , Je Chul Lee , Myung-Je Cho , Woo-Kon Lee , Hyung-Lyun Kang , Jae-Young Song , Seung Chul Baik , Hyeon Su Ro; J. Microbiol. 2016;54(5):387-395. Published online April 20, 2016; DOI: https://doi.org/10.1007/s12275-016-5631-9

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HP0059, an uncharacterized gene of Helicobacter pylori, encodes a 284-aa-long protein containing a nuclear localization sequence (NLS) and multiple leucine-rich heptad repeats. Effects of HP0059 proteins in human stomach cells were assessed by incubation of recombinant HP0059 proteins with the AGS human gastric carcinoma cell line. Wild-type HP0059 proteins showed cytotoxicity in AGS cells in a concentrationdependent manner, whereas NLS mutant protein showed no effect, suggesting that the cytotoxicity is attributed to host nuclear localization. AGS cells transfected with pEGFP-HP0059 plasmid showed strong GFP signal merged to the chromosomal DNA region. The chromosome was fragmented into multiple distinct dots merged with the GFP signal after 12 h of incubation. The chromosome fragmentation was further explored by incubation of AGS chromosomal DNA with recombinant HP0059 proteins, which leaded to complete degradation of the chromosomal DNA. HP0059 protein also degraded circular plasmid DNA without consensus, being an indication of DNase I activity. The DNase was activated by MgCl2, but not by CaCl2. The activity was completely blocked by EDTA. The optimal pH and temperature for DNase activity were 7.0–8.0 and 55°C, respectively. These results indicate that HP0059 possesses a novel DNase I activity along with a role in the genomic instability of human gastric cells, which may result in the transformation of gastric cells.

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Identification of candidate nucleomodulins in ESKAPE bacteria – in silico prediction of bacterial proteins harboring canonical nuclear localization sequences
Laura Riedl, Filip Vilotic, Luzia Burger, Annika Walter, Juergen Fritsch
Frontiers in Cellular and Infection Microbiology.2026;[Epub] CrossRef
Nucleomodulins from gut bacteria: diverse mechanisms of translocation and interaction with host nuclear processes
Sania Korgaonkar, Chandrani Bose, Swadha Anand, Issac Cann
Applied and Environmental Microbiology.2025;[Epub] CrossRef
The Identification of a Novel Nucleomodulin MbovP467 of Mycoplasmopsis bovis and Its Potential Contribution in Pathogenesis
Abdul Raheem, Doukun Lu, Abdul Karim Khalid, Gang Zhao, Yingjie Fu, Yingyu Chen, Xi Chen, Changmin Hu, Jianguo Chen, Huanchun Chen, Aizhen Guo
Cells.2024; 13(7): 604. CrossRef
Molecular Coevolution of Nuclear and Nucleolar Localization Signals inside the Basic Domain of HIV-1 Tat
Margarita A. Kurnaeva, Arthur O. Zalevsky, Eugene A. Arifulin, Olga M. Lisitsyna, Anna V. Tvorogova, Maria Y. Shubina, Gleb P. Bourenkov, Maria A. Tikhomirova, Daria M. Potashnikova, Anastasia I. Kachalova, Yana R. Musinova, Andrey V. Golovin, Yegor S. Va
Journal of Virology.2022;[Epub] CrossRef
Bacterial nucleomodulins and cancer: An unresolved enigma
Abdul Arif Khan, Zakir Khan
Translational Oncology.2021; 14(1): 100922. CrossRef
TatD DNases of African trypanosomes confer resistance to host neutrophil extracellular traps
Kai Zhang, Ning Jiang, Hongyu Chen, Naiwen Zhang, Xiaoyu Sang, Ying Feng, Ran Chen, Qijun Chen
Science China Life Sciences.2021; 64(4): 621. CrossRef
Origin of the nuclear proteome on the basis of pre-existing nuclear localization signals in prokaryotic proteins
Olga M. Lisitsyna, Margarita A. Kurnaeva, Eugene A. Arifulin, Maria Y. Shubina, Yana R. Musinova, Andrey A. Mironov, Eugene V. Sheval
Biology Direct.2020;[Epub] CrossRef

An Easy Way for the Rapid Purification of Recombinant Proteins from Helicobacter pylori Using a Newly Designed Expression Vector: Hyung-Lyun Kang , Jin-Sung Jo , Soon-Uck Kwon , Jae-Young Song , Ji-Hyun Seo , Myung-Je Cho , Seung-Chul Baik , Hee-Shang Youn , Kwang-Ho Rhee , Woo-Kon Lee; J. Microbiol. 2014;52(7):604-608. Published online June 28, 2014; DOI: https://doi.org/10.1007/s12275-014-3679-y

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We constructed a H. pylori expression vector which consisted of both a His-tag and a GST tag as purification tools for recombinant protein and a chloramphenicol resistant cat gene as a reporter. The backbone of the vector pBK contained an ColEI origin of replication and a kanamycin resistant gene. A set of oligos for the His-tag and the PCR product of gst (glutathione S-transferase) gene were inserted sequentially in frame in the multi-cloning site of pBK. The orf of cat was inserted downstream of the gst to generate pBKHGC. The 3' part of H. pylori clpB and flaA were cloned into the vector which was introduced into H. pylori. Recombinant proteins were purified by GSH affinity column, digested with thrombin and were analyzed by western blotting. The final recombinant proteins were successfully purified.

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Gene polymorphisms of pathogenic Helicobacter pylori in patients with different types of gastrointestinal diseases
Yu-Li Chen, Xiao-Qiang Mo, Gan-Rong Huang, Yan-Qiang Huang, Juan Xiao, Li-Juan Zhao, Hong-Yu Wei, Qian Liang
World Journal of Gastroenterology.2016; 22(44): 9718. CrossRef

Profiling of the Bacteria Responsible for Pyogenic Liver Abscess by 16S rRNA Gene Pyrosequencing: Yun Gyu Song , Sang Gun Shim , Kwang Min Kim , Dong-Hae Lee , Dae-Soo Kim , Sang-Haeng Choi , Jae-Young Song , Hyung-Lyun Kang , Seung-Chul Baik , Woo-Kon Lee , Myung-Je Cho , Kwang-Ho Rhee; J. Microbiol. 2014;52(6):504-509. Published online May 29, 2014; DOI: https://doi.org/10.1007/s12275-014-4241-7

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Pyogenic liver abscess (PLA) is a severe disease with considerable mortality and is often polymicrobial. Understanding the pathogens that cause PLA is the basis for PLA treatment. Here, we profiled the bacterial composition in PLA fluid by pyrosequencing the 16S ribosomal RNA (rRNA) gene based on next-generation sequencing (NGS) technology to identify etiological agents of PLA and to provide information of their 16S rRNA sequences for application to DNA-based techniques in the hospital. Twenty patients with PLA who underwent percutaneous catheter drainage, abscess culture, and blood culture for isolates were included. Genomic DNAs from abscess fluids were subjected to polymerase chain reaction and pyrosequencing of the 16S rRNA gene with a 454 GS Junior System. The abscess and blood cultures were positive in nine (45%) and four (20%) patients, respectively. Pyrosequencing of 16S rRNA gene showed that 90% of the PLA fluid samples contained single or multiple genera of known bacteria such as Klebsiella, Fusobacterium, Streptococcus, Bacteroides, Prevotella, Peptostreptococcus, unassigned Enterobacteriaceae, and Dialister. Klebsiella was predominantly found in the PLA fluid samples. All samples that carried unassigned bacteria had 26.8% reads on average. We demonstrated that the occurrence of PLA was associated with eight known bacterial genera as well as unassigned bacteria and that 16S rRNA gene sequencing was more useful than conventional culture methods for accurate identification of bacterial pathogens from PLA.

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Genetic Organization and Conjugal Plasmid DNA Transfer of pHP69, a Plasmid from a Korean Isolate of Helicobacter pylori: Jung-Soo Joo , Jae-Young Song , Seung-Chul Baik , Woo-Kon Lee , Myung-Je Cho , Kon-Ho Lee , Hee-Shang Youn , Ji-Hyun Seo , Kwang-Ho Rhee , Hyung-Lyun Kang; J. Microbiol. 2013;51(6):889-889.; DOI: https://doi.org/10.1007/s12275-013-0727-y

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Abstract PDF: In the article by Joo et al. published in the Journal of Microbiology 2012; 50, 955-961. Dr. YoungAh Cho, one of coauthor’s name was inadvertently missing. Complete authors information is as follow. Jung-Soo Joo4, Jae-Young Song1,3, Seung-Chul Baik1,3, Woo-Kon Lee1,3, Myung-Je Cho1,3, Kon-Ho Lee1,3, YoungAh Cho1, Hee-Shang Youn2, Ji-Hyun Seo2, Kwang-Ho Rhee1,3, and Hyung-Lyun Kang1,3*

Genetic Organization and Conjugal Plasmid DNA Transfer of pHP69, a Plasmid from a Korean Isolate of Helicobacter pylori: Jung-Soo Joo , Jae-Young Song , Seung-Chul Baik , Woo-Kon Lee , Myung-Je Cho , Kon-Ho Lee , Hee-Shang Youn , Ji-Hyun Seo , Kwang-Ho Rhee , Hyung-Lyun Kang; J. Microbiol. 2012;50(6):955-961. Published online December 30, 2012; DOI: https://doi.org/10.1007/s12275-012-2580-9

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We isolated pHP69, a 9,153 bp plasmid from Helicobacter pylori with a 33.98% (G+C) content. We identified 11 open reading frames (ORFs), including replication initiation protein A (repA), fic (cAMP-induced filamentation protein), mccC, mccB, mobA, mobD, mobB, and mobC, as well as four 22 bp tandem repeat sequences. The nucleic acid and predicted amino acid sequences of these ORFs exhibited significant homology to those of other H. pylori plasmids. pHP69 repA encodes a replication initiation protein and its amino acid sequence is similar to those of replicase proteins from theta-type plasmids. pHP69 contains two types of repeat sequences (R1 and R2), a MOBHEN family mobilization region comprising mobC, mobA, mobB, and mobD, and genes encoding microcin B and C. Among the 36 H. pylori strains containing plasmids, mobA or mccBC are present in 12 or 6, respectively and 3 contain both genes. To examine intrinsic capability of H. pylori for conjugative plasmid transfer, a shuttle vector pBHP69KH containing pHP69 and replication origin of pBR322 was constructed. It was shown that this vector could stably replicate and be mobilized among clinical H. pylori strains and demonstrated to gene transfer by natural plasmid.

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Helicobacter pylori γ-Glutamyltranspeptidase Induces Cell Cycle Arrest at the G1-S Phase Transition: Kyung-Mi Kim , Seung-Gyu Lee , Jung-Min Kim , Do-Su Kim , Jea-Young Song , Hyung-Lyun Kang , Woo-Kon Lee , Myung-Je Cho , Kwang-Ho Rhee , Hee-Shang Youn , Seung-Chul Baik; J. Microbiol. 2010;48(3):372-377. Published online June 23, 2010; DOI: https://doi.org/10.1007/s12275-010-9293-8

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Abstract PDF

In our previous study, we showed that Helicobacter pylori γ-glutamyltranspeptidase (GGT) is associated with H. pylori-induced apoptosis through a mitochondrial pathway. To better understand the role of GGT in apoptosis, we examined the effect of GGT on cell cycle regulation in AGS cells. To determine the effect of recombinant GGT (rGGT) on cell cycle distribution and apoptosis, rGGT-treated and untreated AGS cells were analyzed in parallel by flow cytometry using propidium iodide (PI). We found that rGGT inhibited the growth of AGS cells in a time-dependent manner, and that the pre-exposure of cells to a caspase-3 inhibitor (z-DEVD-fmk) effectively blocked GGT-induced apoptosis. Cell cycle analysis showed G1 phase arrest and apoptosis in AGS cells following rGGT treatment. The rGGT-mediated G1 phase arrest was found to be associated with down-regulation of cyclin E, cyclin A, Cdk 4, and Cdk 6, and the up-regulation of the cyclindependent kinase (Cdk) inhibitors p27 and p21. Our results suggest that H. pylori GGT induces cell cycle arrest at the G1-S phase transition.

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