• 생명과학회지
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• 제27권4호
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• pp.482-499
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• 2017

Allergy 질환은 지난 십여년 동안 개발도상국을 포함해서 전 세계적으로 증가하고 있다. Allergy 염증 반응은 수지상 세포와 같은 항원제시 세포에 의한 allergy 항원섭취를 시작으로 하여 Th2 면역 반응에 의해서 일어난다. 장내 미생물은 신체의 대사나 생리적 기능을 조절하고, 생애 초기의 면역 체계 성숙과 일생 동안 면역 체계 항상성 및 상피세포 총체성에 기여한다. Bifidobacteria는 Th1/Th2 balance에 strain-specific 한 면역 자극 특성을 가지며, TSLP와 IgE 발현을 억제 시키고 Flg과 FoxP3 발현을 촉진 시켜 allergy를 완화시킨다. 또한 Unmethylated CpG motif ODN은 B 세포와 수지상 세포의 TLR9에 의해 인식 되어 선천성과 적응성 면역 반응을 유도하고, Clostridium butyricum에 의해서 생산된 butyrate는 수지상 세포의 anti-inflammatory 유전자의 발현을 유도하기 위해 GPR109a signaling pathway를 활성화시키고, GPR43 활성화를 통하여 tTreg 세포 proliferation을 직접 자극하거나 HADC 활성을 억제시켜 Foxp3 gene intronic enhancer의 histone H3 acetylation을 통해 naive $CD4^+$ T 세포를 pTreg 세포로 분화시킨다.

• BMB Reports
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• 제54권8호
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• pp.413-418
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• 2021

ErbB3-binding protein 1 (EBP1) is a multifunctional protein associated with neural development. Loss of Ebp1 leads to upregulation of the gene silencing unit suppressor of variegation 3-9 homolog 1 (Suv39H1)/DNA (cytosine 5)-methyltransferase (DNMT1). EBP1 directly binds to the promoter region of DNMT1, repressing DNA methylation, and hence, promoting neural development. In the current study, we showed that EBP1 suppresses histone methyltransferase activity of Suv39H1 by promoting ubiquitin-proteasome system (UPS)-dependent degradation of Suv39H1. In addition, we showed that EBP1 directly interacts with Suv39H1, and this interaction is required for recruiting the E3 ligase MDM2 for Suv39H1 degradation. Thus, our findings suggest that EBP1 regulates UPS-dependent degradation of Suv39H1 to govern proper heterochromatin assembly during neural development.

• Asian-Australasian Journal of Animal Sciences
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• 제33권6호
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• pp.1023-1033
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• 2020

Objective: The efficiency of the knock-in process is very important to successful gene editing in domestic animals. Recently, it was reported that transient loosening of the nucleosomal folding of transcriptionally inactive chromatin might have the potential to enhance homologous recombination efficiency. The objective of this study was to determine whether histone deacetylases (HDAC) inhibitor and RAD51 recombinase (RAD51) expression were associated with increased knock-in efficiency on the β-casein (bCSN2) gene locus in mammary alveolar-large T antigen (MAC-T) cells using the transcription activator-like effector nucleases (TALEN) system. Methods: MAC-T cells were treated with HDAC inhibitors, valproic acid, trichostatin A, or sodium butyrate for 24 h, then transfected with a knock-in vector, RAD51 expression vector and TALEN to target the bCSN2 gene. After 3 days of transfection, the knock-in efficiency was confirmed by polymerase chain reaction and DNA sequencing of the target site. Results: The level of HDAC 2 protein in MAC-T cells was decreased by treatment with HDAC inhibitors. The knock-in efficiency in MAC-T cells treated with HDAC inhibitors was higher than in cells not treated with inhibitors. However, the length of the homologous arm of the knock-in vector made no difference in the knock-in efficiency. Furthermore, DNA sequencing confirmed that the precision of the knock-in was more efficient in MAC-T cells treated with sodium butyrate. Conclusion: These results indicate that chromatin modification by HDAC inhibition and RAD51 expression enhanced the homologous recombination efficiency on the bCSN2 gene locus in MAC-T cells.

• Biomolecules & Therapeutics
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• 제32권2호
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• pp.214-223
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• 2024

Metabolic abnormalities in the liver are closely associated with diverse metabolic diseases such as non-alcoholic fatty liver disease, type 2 diabetes, and obesity. The aim of this study was to evaluate the ameliorating effect of robinetin (RBN) on the significant pathogenic features of metabolic failure in the liver and to identify the underlying molecular mechanism. RBN significantly decreased triglyceride (TG) accumulation by downregulating lipogenesis-related transcription factors in AML-12 murine hepatocyte cell line. In addition, mice fed with Western diet (WD) containing 0.025% or 0.05% RBN showed reduced liver mass and lipid droplet size, as well as improved plasma insulin levels and homeostatic model assessment of insulin resistance (HOMA-IR) values. CD38 was identified as a target of RBN using the BioAssay database, and its expression was increased in OPA-treated AML-12 cells and liver tissues of WD-fed mice. Furthermore, RBN elicited these effects through its anti-histone acetyltransferase (HAT) activity. Computational simulation revealed that RBN can dock into the HAT domain pocket of p300, a histone acetyltransferase, which leads to the abrogation of its catalytic activity. Additionally, knock-down of p300 using siRNA reduced CD38 expression. The chromatin immunoprecipitation (ChIP) assay showed that p300 occupancy on the promoter region of CD38 was significantly decreased, and H3K9 acetylation levels were diminished in lipid-accumulated AML-12 cells treated with RBN. RBN improves the pathogenic features of metabolic failure by suppressing the p300-CD38 axis through its anti-HAT activity, which suggests that RBN can be used as a new phytoceutical candidate for preventing or improving this condition.

• 생명과학회지
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• 제29권9호
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• pp.1002-1009
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• 2019

생애 초기 유해 경험은 우울증의 위험성을 높이며, 성인기 스트레스의 민감성에 영향을 미칠 수 있다. 출생 후 모성 분리(MS)로 인한 성인기 스트레스(장기간 예측 불가능한 스트레스; CUS)의 취약성에 p11 유전자의 후성유전기전이 영향을 미치는 지를 확인하였다. 출생 직후부터 21일 동안 하루 3시간 동안 새끼 생쥐를 어미 생쥐로부터 분리시켜 새끼 생쥐가 성체가 되었을 때 CUS를 3주 동안 매일 적용하였다. Real time PCR기법으로 해마의 p11 발현 양을 측정하였고, 염색질 면역 침전 분석법으로 p11 promoter의 히스톤 H3 아세틸화 및 메틸화 양을 측정하였다. 강제수영검사에서 우울 유사 행동을 측정하였다. MS군 및 CUS군은 p11 mRNA 발현 양을 유의하게 감소시켰으며, MS+CUS군은 CUS군에 비해 p11 발현 양을 유의하게 증가시켰다. 또한 MS+CUS군은 CUS군에 비해 H3 아세틸화를 감소시켰다. 이러한 감소는 HDAC5 mRNA 발현 증가와 일치하였다. MS+CUS군은 CUS군에 비해 H3K4 메틸화를 감소시켰으며, H3K27 메틸화를 증가시켰다. 강제수영검사에서 p11 발현이 가장 많이 감소된 MS+CUS군이 대조군에 비해 더 긴 부동 시간을 나타내었다. 출생 후 모성 분리를 경험하고 성체 기간에 스트레스를 함께 받은 생쥐는 성체기간에만 스트레스를 받은 생쥐보다 훨씬 더 큰 후성유전 변화를 보여주었다. 생애 초기 유해 경험은 해마에서 p11 유전자의 히스톤 변형을 통해 성체 스트레스 효과를 더 악화시키는 것으로 생각된다.

• Molecules and Cells
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• 제42권1호
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• pp.17-27
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• 2019

Ubiquitin-specific protease 44 (USP44) has been implicated in tumor progression and metastasis across various tumors. However, the function of USP44 in prostate cancers and regulatory mechanism of histone-modifying enzymes by USP44 in tumors is not well-understood. Here, we found that enhancer of zeste homolog 2 (EZH2), a histone H3 lysine 27 methyltransferase, is regulated by USP44. We showed that EZH2 is a novel target of USP44 and that the protein stability of EZH2 is upregulated by USP44-mediated deubiquitination. In USP44 knockdown prostate cancer cells, the EZH2 protein level and its gene silencing activity were decreased. Furthermore, USP44 knockdown inhibited the tumorigenic characteristics and cancer stem cell-like behaviors of prostate cancer cells. Inhibition of tumorigenesis caused by USP44 knockdown was recovered by ectopic introduction of EZH2. Additionally, USP44 regulates the protein stability of oncogenic EZH2 mutants. Taken together, our results suggest that USP44 promotes the tumorigenesis of prostate cancer cells partly by stabilizing EZH2 and that USP44 is a viable therapeutic target for treating EZH2-dependent cancers.

• 아시안잔디학회지
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• 제24권2호
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• pp.165-169
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• 2010

Fusarium circinatum은 소나무류에 푸사리움 가지마름병을 유발하는 병원균으로 알려져 있다. 본 연구에서는 골프장의 리기다소나무로부터 병원균을 분리하였으며, 이들의 형태적인 특성을 관찰하고 분자생물학적인 방법을 이용하여 동정한 결과 F. circinatum 임을 확인하였다. 특히 histone H3 유전자와 IGS rDNA의 염기서열을 분석한 결과 분리된 균들의 염기서열은 동일하였으며, 유성생식(mating)에 관여하는 유전자를 분석한 결과 모두 MAT1-1 유형임을 확인하였다. 또한 푸사리움 가지마름병에 대한 감수성 수종인 리기다소나무를 비롯하여 저항성 수종인 잣나무와 소나무의 메탄올 추출물을 만들어 이들의 병원균에 대한 항균력을 검정하였다. 그 결과 저항성 수종인 잣나무와 소나무의 추출물에서 리기다소나무 보다 높은 항균력을 확인 하였다.

• Biomolecules & Therapeutics
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• 제27권6호
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• pp.530-539
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• 2019

Brain aging is an inevitable process characterized by structural and functional changes and is a major risk factor for neurodegenerative diseases. Most brain aging studies are focused on neurons and less on astrocytes which are the most abundant cells in the brain known to be in charge of various functions including the maintenance of brain physical formation, ion homeostasis, and secretion of various extracellular matrix proteins. Altered mitochondrial dynamics, defective mitophagy or mitochondrial damages are causative factors of mitochondrial dysfunction, which is linked to age-related disorders. Etoposide is an anti-cancer reagent which can induce DNA stress and cellular senescence of cancer cell lines. In this study, we investigated whether etoposide induces senescence and functional alterations in cultured rat astrocytes. Senescence-associated ${\beta}$-galactosidase (SA-${\beta}$-gal) activity was used as a cellular senescence marker. The results indicated that etoposide-treated astrocytes showed cellular senescence phenotypes including increased SA-${\beta}$-gal-positive cells number, increased nuclear size and increased senescence-associated secretory phenotypes (SASP) such as IL-6. We also observed a decreased expression of cell cycle markers, including PhosphoHistone H3/Histone H3 and CDK2, and dysregulation of cellular functions based on wound-healing, neuronal protection, and phagocytosis assays. Finally, mitochondrial dysfunction was noted through the determination of mitochondrial membrane potential using tetramethylrhodamine methyl ester (TMRM) and the measurement of mitochondrial oxygen consumption rate (OCR). These data suggest that etoposide can induce cellular senescence and mitochondrial dysfunction in astrocytes which may have implications in brain aging and neurodegenerative conditions.

• Journal of Ginseng Research
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• 제43권1호
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• pp.1-9
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• 2019

Background: Korean ginseng is an important cash crop in Asian countries. However, plant yield is reduced by pathogens. Among the Ilyonectria radicicola-species complex, I. mors-panacis is responsible for root-rot and replant failure of ginseng in Asia. The development of new methods to reveal the existence of the pathogen before cultivation is started is essential. Therefore, a quantitative real-time polymerase chain reaction method was developed to detect and quantify the pathogen in ginseng soils. Methods: In this study, a species-specific histone H3 primer set was developed for the quantification of I. mors-panacis. The primer set was used on DNA from other microbes to evaluate its sensitivity and selectivity for I. mors-panacis DNA. Sterilized soil samples artificially infected with the pathogen at different concentrations were used to evaluate the ability of the primer set to detect the pathogen population in the soil DNA. Finally, the pathogen was quantified in many natural soil samples. Results: The designed primer set was found to be sensitive and selective for I. mors-panacis DNA. In artificially infected sterilized soil samples, using quantitative real-time polymerase chain reaction the estimated amount of template was positively correlated with the pathogen concentration in soil samples ($R^2=0.95$), disease severity index ($R^2=0.99$), and colony-forming units ($R^2=0.87$). In natural soils, the pathogen was recorded in most fields producing bad yields at a range of $5.82{\pm}2.35pg/g$ to $892.34{\pm}103.70pg/g$ of soil. Conclusion: According to these results, the proposed primer set is applicable for estimating soil quality before ginseng cultivation. This will contribute to disease management and crop protection in the future.

• 한국환경성돌연변이발암원학회지
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• 제24권4호
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• pp.171-179
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• 2004

Cytochrome P4503A4(CYP3A4) is the most abundnat CYPs in human liver, comparising approximately 30% of the total liver CYPs contents ans is involbed in the metabolism of more than 60% of currently used therapeutic drugs. The expression of CYP3A4 is induced by a variety of structurally unrelated xonobiotics including the antibiotic rifampicin and endogenous hormones, and might be mediated through steroid and xenobiotic receptor(SXR) system. The molecular mechanisms underlying regulation of CYP3A4 gene expression hae not been understood. In order to gain the insight of the molecular mechanism of CYP3A4 gene expression, study has been undertaken to investigate if the histone deacelylation is involved in the regulation of CYP3A4 gene expression by proximal promoter or not. Also SXR was investigated to see if they were involved in the regulation of CYP3A4 proximal promoter activity. HepG2 or Hena-I cells were transfected with a plasmid containing~1kb of the CYP3A4 proximal promoter region (-863 to +64bp) cloned in front of a reporter gene, luciferase, in the presence or absence of SXR or hER. Transfected cells were treated with CYP3A4 inducers such as rifampicin, PCN and RU 486, or with estradiol, in order to exmine to regulation of CYP3A4 gene expression in the presence or absence of trichostatin A (TSA). In HepG2 cells, CYP3A4 inducers and estradiol increased significantly the luciferase activity by CYP3A4 proximal promoter, only when TSA was co-treated after SXR cotransfection. In the case of Hepa-I cells CYP3A4 inducers and estradiol incressed modestly the luciferase activity when TSA was co-treated, but this increment was not enhanced by SXR cotransfection in contrast to HepG2 cells. Taken together, these results indicated that the inhibition of histone deacetylation was required to SXR-mediated increase in CYP3A4 proximal promoter region when rifampicin, or PCN was treated. Futher a trans-activation by SXR may demand other species-specific transcription factors.