• Title/Summary/Keyword: H-NS

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H-NS binding on dicA promoter DNA inhibits dicA gene expression (dicA promoter DNA에 붙는 H-NS 단백질에 의한 dicA 유전자의 발현 조절)

  • Yun, Sang Hoon;Lee, Yonho;Lim, Heon M.
    • Korean Journal of Microbiology
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    • v.55 no.3
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    • pp.191-198
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    • 2019
  • H-NS binds to promoter DNA and works as a general transcription silencer. DicA protein, by binding to the promoter DNA of dicA, activates dicA expression and at the same time inhibits expression of dicF and dicB, thus, exerting cell division control in Escherichia coli. H-NS complexed with a nucleoid protein Cnu was known to be involved in dicA expression. However, the exact nature of H-NS binding to dicA promoter DNA and the consequences of H-NS binding in expression of dicA is not clear. In this study, we explored the DNA binding activity of H-NS on the promoter DNA of dicA and found that H-NS binding occurs exclusively to the dicA promoter DNA. We never observed, however, H-NS binding at the vicinity of the dicA promoter. Temperature dependent oligomerization of H-NS was observed during DNA binding and the Cnu protein enhances the oligomerization process of H-NS binding. In vivo measurement of dicA expression in an hns deleted strain showed that dicA expression increased. These results demonstrated that H-NS binds specifically to dicA promoter DNA and functions as a transcription silencer.

H-NS Silences Gene Expression of LeuO, the Master Regulator of the Cyclic(Phe-Pro)-dependent Signal Pathway, in Vibrio vulnificus

  • Park, Na-Young;Lee, Keun-Woo;Kim, Kun-Soo
    • Journal of Microbiology and Biotechnology
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    • v.30 no.6
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    • pp.830-838
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    • 2020
  • The histone-like nucleoid structuring protein (H-NS) is an abundant global regulator of environmentally controlled gene expression. Herein, we demonstrate that H-NS represses the expression of LeuO, the master regulator of the cyclic(Phe-Pro)-dependent signaling pathway, by directly binding to the upstream region of the gene. H-NS binds to a long stretched region (more than 160-bp long), which overlaps with binding sites for ToxR and LeuO. A high quantity of H-NS outcompetes ToxR for binding to the cis-acting element of leuO. However, our footprinting analyses suggests that the binding of H-NS is relatively weaker than LeuO or ToxR at the same molarity. Considering that the DNA nucleotide sequences of the upstream regions of leuO genes are highly conserved among various Vibrio, such patterns as those found in V. vulnificus would be a common feature in the regulation of leuO gene expression in Vibrionaceae. Taken together, these results suggest that, in species belonging to Vibrionaceae, H-NS regulates the expression of leuO as a basal stopper when cFP-ToxR mediated signaling is absent.

Regulation of gene expression by histone-like proteins in bacteria (박테리아의 히스톤 유사 단백질에 의한 유전자 발현 조절)

  • Park, Shinae;Lee, Jung-Shin
    • Korean Journal of Microbiology
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    • v.54 no.1
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    • pp.1-8
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    • 2018
  • A prokaryotic cell has various histone-like proteins also known as nucleoid-associated proteins (NAPs). These proteins bind AT-rich sequence at DNA, which induce DNA wrapping, bending, and bridging, and subsequently regulate the gene expression in bacteria. Because NAPs function in transcriptional silencing of virulence genes, it is important to study their roles in gene silencing and specific mechanisms of these proteins. In this review, we discussed two well-known NAPs, H-NS, and HU, and summarized their roles for gene expression in Escherichia coli and Salmonella Typhimurium. Through the oligomerization and filamentation of H-NS, it represses the expression of virulence genes in human pathogenic bacteria, such as Salmonella Typhimurium, and it works with other NAPs positively or negatively. Recently, H-NS also regulates typhoid toxin expression, which causes typhoid fever and systemic disease in human. Additionally, HU regulates the expression of genes related to both virulence and physiology of Salmonella. Therefore, we suggest that NAPs like H-NS and HU are crucial factors to reveal the molecular mechanisms of virulence gene expression in bacteria.

Cupric Complexes Produced from the Reaction of Cupric Nitrate Trihydrate with S-2-Pyridyl Thioates

  • Choi, Young-Nam;So, Hyun-Soo;Lee, Jae-In;Kim, Sung-Gak
    • Bulletin of the Korean Chemical Society
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    • v.7 no.5
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    • pp.385-388
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    • 1986
  • The reaction of cupric nitrate trihydrate with S-2-pyridyl thioates in acetonitrile was studied. The major products were the corresponding carboxylic acids and $[Cu(NO_3)(C_5H_4NS)(C_5H_5NS$)] (Complex A). Sometimes $[Cu(NO_3)(C_5H_4NS)(H_2O$)] was also obtained in addition to Complex A. When Complex A was recrystallized in dimethylsulfoxide, $[Cu(NO_3)(C_5H_4NS)(C_5H_5NS)$ {$(CH_3)_2SO$}$_2]{\cdot}2H_2O$ was crystallized. The structures of these copper complexes and the role of cupric nitrate in the hydrolysis of S-2-pyridyl thioates are discussed.

The Ca2+-activated K+ (BK) Channel-opener NS 1619 Prevents Hydrogen Peroxide-induced Cell Death and Mitochondrial Dysfunction in Retinal Pigment Epithelial Cells (망막 색소상피세포에서 산화성 세포 손상과 미토콘드리아기능 저해에 미치는 NS 1619의 보호 효과)

  • Kang, Jae Hoon;Woo, Jae Suk
    • Journal of Life Science
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    • v.27 no.11
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    • pp.1349-1356
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    • 2017
  • Potassium channel openers (KCOs) produce physiological and pharmacological defense mechanisms against cell injuries caused by oxidative stress of diverse origins. Openings of mitochondrial and plasmalemmal $K^+$ channels are involved in the defense mechanisms. This study tested whether NS 1619, an opener of large-conductance BK channels, has a similar beneficial influence on the pigment epithelial cells of retinas. The human retinal pigment epithelial cell line ARPE-19 was exposed to $H_2O_2$-induced oxidative stress in the absence and presence of NS 1619. The degrees of the cells' injuries were assessed by analyzing the cells' trypan-blue exclusion abilities and TUNEL staining. NS 1619 produced remarkable protections against cell injuries caused by $H_2O_2$. It prevented apoptotic and necrotic cell deaths. The protective effect of NS 1619 was significantly diminished when the cells were treated with NS 1619 in combination with the BK channel-blocker paxilline. NS 1619 significantly ameliorated cellular ATP deprivations in $H_2O_2$-treated cells. It helped mitochondria preserve their functional integrity, which was estimated by their MTT reduction abilities and mitochondrial membrane potential. In conclusion, it was suggested that NS 1619 had a beneficial effect on mitochondria in regards to preserving their functional integrity under oxidative stress, and it produces defense mechanisms against oxidant-induced cell injuries in ARPE-19 cells.

The Fluorescence Behavior of the Responsive Macrocycle by Aromatic Imine Molecules

  • Choi, Chang-Shik;Jeon, Ki-Seok;Lee, Ki-Hwan
    • Journal of Photoscience
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    • v.11 no.32
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    • pp.71-74
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    • 2004
  • The macrocycle L exhibited a switch on-off behavior through the fluorescent responses by aromatic imine molecule 1 (X=H) / trifluoroacetic acid (TFA). In the 'switch on' state, it was supposed that the aromatic imine molecule 1 is in the cavity of macrocycle L and a photoinduced electron transfer (PET) from the nitrogen of azacrown part to the anthryl group is inhibited by the interaction between the aromatic imine molecule 1 and the azacrown part of macrocycle L. In the 'switch off' state, it was supposed that the protonated imine molecule 1 is induced by the continuous addition of TFA and a repulsion between the protonated azacrown part and the protonated imine molecule 1 is occurred. It was considered that this process induces the intermolecular PET from the protonated imine molecule 1 to the anthryl group of macrocycle L because of a proximity effect between the anthryl group and the protonated imine molecule 1. From the investigation of the transient emission decay curve, the macrocycle L showed three components (3.45 ns (79.72%), 0.61 ns (14.53%), and 0.10 ns (5.75%). When the imine molecule 1 was added in the macrocycle L as molar ratio=1:1, the first main component showed a little longer lifetime as 3.68 ns (82.75%) although the other two components were similar as 0.64 ns (14.28%) and 0.08 ns (2.96%). On the contrary, when the imine molecule 3 (X=C1) was added in the macrocycle L as molar ratio=l:1, all the three components were decreased such as 3.27 ns (69.83%), 0.44 ns (13.24%), and 0.06 ns (16.93%). The fluorescent pH titration of macrocycle L was carried out from pH=3 to pH=9. The macrocycle L and C $U^{2+}$- macrocycle L complex were intersected at about pH=5, while the E $u^{3+}$ -macrocycle L complex was intersected at about pH=5.5. In addtion, we investigated the fluorescence change of macrocycle L as a function of the substituent constant ($\sigma$$_{p}$$^{o}$) showing in the para-substituent with electron withdrawing groups (X=F, Cl) and electron donating groups (X=C $H_3$, OC $H_3$, N(C $H_3$)$_2$), respectively, as well as non-substituent (X=H).).ctively, as well as non-substituent (X=H).

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Molecular and Structural Characterization of the Domain 2 of Hepatitis C Virus Non-structural Protein 5A

  • Liang, Yu;Kang, Cong Bao;Yoon, Ho Sup
    • Molecules and Cells
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    • v.22 no.1
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    • pp.13-20
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    • 2006
  • Hepatitis C virus (HCV) non-structural protein 5A protein (NS5A), which consists of three functional domains, is involved in regulating viral replication, interferon resistance, and apoptosis. Recently, the three-dimensional structure of the domain 1 was determined. However, currently the molecular basis for the domains 2 and 3 of HCV NS5A is yet to be defined. Toward this end, we expressed, purified the domain 2 of the NS5A (NS5A-D2), and then performed biochemical and structural studies. The purified domain 2 was active and was able to bind NS5B and PKR, biological partners of NS5A. The results from gel filtration, CD analysis, 1D $^1H$ NMR and 2D $^1H-^{15}N$ heteronuclear single quantum correlation (HSQC) spectroscopy indicate that the domain 2 of NS5A appears to be flexible and disordered.

Effect of Morphology and Dispersibility of Silica Nanoparticles on the Mechanical Behaviour of Cement Mortar

  • Singh, Lok Pratap;Goel, Anjali;Bhattachharyya, Sriman Kumar;Ahalawat, Saurabh;Sharma, Usha;Mishra, Geetika
    • International Journal of Concrete Structures and Materials
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    • v.9 no.2
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    • pp.207-217
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    • 2015
  • The influence of powdered and colloidal nano-silica (NS) on the mechanical properties of cement mortar has been investigated. Powdered-NS (~40 nm) was synthesized by employing the sol-gel method and compared with commercially available colloidal NS (~20 nm). SEM and XRD studies revealed that the powdered-NS is non-agglomerated and amorphous, while colloidal-NS is agglomerated in nature. Further, these nanoparticles were incorporated into cement mortar for evaluating compressive strength, gel/space ratio, portlandite quantification, C-S-H quantification and chloride diffusion. Approximately, 27 and 37 % enhancement in compressive strength was observed using colloidal and powdered-NS, respectively, whereas the same was up to 19 % only when silica fume was used. Gel/space ratio was also determined on the basis of degree of hydration of cement mortar and it increases linearly with the compressive strength. Furthermore, DTG results revealed that lime consumption capacity of powdered-NS is significantly higher than colloidal-NS, which results in the formation of additional calcium-silicate-hydrate (C-S-H). Chloride penetration studies revealed that the powdered-NS significantly reduces the ingress of chloride ion as the microstructure is considerably improved by incorporating into cement mortar.

Inefficient Transmissibility of NS-Truncated H3N8 Equine Influenza Virus in Dogs

  • Na, Woonsung;Song, Manki;Yeom, Minjoo;Park, Nanuri;Kang, Bokyu;Moon, Hyoungjoon;Jeong, Dae-Gwin;Kim, Jeong-Ki;Song, Daesub
    • Journal of Microbiology and Biotechnology
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    • v.25 no.3
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    • pp.317-320
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    • 2015
  • H3N8 equine influenza virus (EIV) causes respiratory diseases in the horse population, and it has been demonstrated that EIV can transmit into dogs owing to its availability on receptors of canine respiratory epithelial cells. Recently, we isolated H3N8 EIV from an EIV-vaccinated horse that showed symptoms of respiratory disease, and which has a partially truncated nonstructural gene (NS). However, it is not clear that the NS-truncated EIV has an ability to cross the host species barrier from horses to dogs as well. Here, we experimentally infected the NS-truncated H3N8 EIV into dogs, and monitored their clinical signs and viral load in respiratory organs to determine the virus's transmissibility.

Microarray analysis of gene expression profile by treatment of Nelumbinis Semen in LPS activated BV-2 microglial cells (LPS로 자극한 BV-2 microglial cell에서 Microarray를 통한 련자육(蓮子肉)의 유전자 발현 분석)

  • Kim, Soo-Oh;Lim, Byung-Mook;Bae, Hyun-Su
    • Korean Journal of Oriental Medicine
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    • v.14 no.1
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    • pp.145-160
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    • 2008
  • Nelumbinis Semen(NS) has been used in traditional medicine to treat diseases such as depression and diarrhea. In inflammatory responses, microglia produces molecules which are known to play roles in the central nervous system. And we previously studied NS inhibited nitric oxide synthase and secretion of tumor necrosis factor alpha. To explore the global gene expression profiles in BV-2 microglial cell line treated with NS, microarray analysis was performed. The cells were treated with LPS or NS plus LPS for 30min, Ih, 3h, and 6h, respectively. Of 45,101 known genes, with cutoff value of 3-fold change in the expression, 340, 644, 280 and 219 genes were upregulated and 503, 570, 694 and 484 were downregulated in NS treated cells at each time point. The results of the present study shows that treatment of NS reversed the LPS-induced upregulation of such genes as Ecoxsackievirus and adenovirus receptor(CAR), pellino 1, and S100P binding protein. It is thought that microarrays will play an ever-growing role in the advance of our understanding of the pharmacologic actions NS.

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