• 미생물학회지
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• 제45권4호
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• pp.312-317
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• 2009

사상성 모델 진균인 Aspergillus nidulans에 존재하는 forkhead 유전자들의 구조와 기능을 체계적으로 연구하기 위해 유전체 분석을 통해 forkhead 도메인을 가지고 있는 6개의 forkhead 유전자를 발견하였다. 이들 중 4개의 유전자들은 다른 진균에서 발견되는 forkhead 유전자들과 전반적인 유사성이 있었으나, 2개의 유전자들은 다른 종에서는 보존되어있지 않은 A. nidulans 특이적인 유전자들이었다. A. nidulans 특이적 forkhead 유전자들 중 하나인 fkhF(AN8949.2) 유전자는 염색체 7번에 위치하고 있고, ORF는 2,337 bp로 구성되어 있으며 778개의 아미노산을 암호화하고 있는 것으로 추정되었다. 예상되는 FkhF 단백질의 N-말단 부위에 보존된 forkhead 도메인을 가지고 있었으며, 이 유전자의 기능을 분석하기 위해 유전자제거 돌연변이 균주를 제조하였다. fkhF 유전자 결손돌연변이주는 유성분화 유도 조건 등을 포함한 여러 고체배지 배양 조건에서 유성분화에는 영향을 미치지 않았으나 무성포자병(conidiophore)의 생성 밀도와 성숙도에 영향을 주는 것으로 관찰되었고, 야생형과 달리 진탕배양시에 무성포자병을 형성함이 관찰되었다. 이는 fkhF 유전자가 A. nidulans의 부적절한 무성분화를 억제하고 정상적인 무성포자 성숙과정에 관여하는 유전자라는 것을 보여준다.

• 한국균학회지
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• 제38권2호
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• pp.160-166
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• 2010

모델 사상성 진균 Aspergillus nidulans는 분화과정을 연구하는 진핵세포 시스템으로 사용되어 왔다. 이러한 분화과정은 매우 다양한 유전자들의 발현을 통하여 조절되며 이와 관련된 다양한 전사요소들의 기능이 연구되어 왔다. 이들 중 forkhead 유전자는 일반적으로 감수분열 및 세포주기 조절에 중요한 역할을 하는 것으로 알려져 왔으며 A. nidulans에서도 유사한 기능을 하리라 예상되어 왔다. 이와 관련된 연구를 위하여 A. nidulans 유전체에 존재하는 6개의 forkhead 유전자를 발견, 확보하였고, 최근에는 효모 및 다른 진균에서는 발견되지 않는 A. nidulans 특이적 forkhead 유전자인 fkhF의 구조와 기능이 분석된 바 있다. 본 연구에서는 fkhF와 매우 유사한 단백질 서열을 가지고 있는 fkhE(AN2025.3) 유전자의 기능을 분석하였다. 본 유전자의 기능을 분석하기 위해 RT-PCR을 통하여 cDNA 서열을 분석한 결과 약 3종류의 서로 다른 mRNA가 존재하는 것이 밝혀졌고 이는 alternative splicing에 의한 것으로 추정되었다. 이들 3종류의 mRNA중 한 종류만 정상적인 ORF를 가지고 있으며 조사한 전체 cDNA 발현의 61%를 차지하였다. fkhE 유전자는 718개의 아미노산을 암호화하는 하나의 ORF를 가지고 있었으며 N 말단에 보존된 forkhead 도메인을 가지고 있었다. fkhE 유전자를 제거한 유전자 제거 돌연변이 균주는 fkhF와 유사하게 고체배지에서는 무성포자의 형성이 저해되었으나 유성분화에는 별다른 영향을 미치지 않았으며 액체 진탕배양에서는 야생형과 다르게 무성포자병(conidiophore)이 형성되었다. 이러한 결과는 fkhE 유전자가 무성분화에 관련되었음을 보여준다.

• Journal of Microbiology and Biotechnology
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• 제33권11호
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• pp.1420-1427
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• 2023

The forkhead domain genes are important for development and morphogenesis in fungi. Six forkhead genes fkhA-fkhF have been found in the genome of the model filamentous Ascomycete Aspergillus nidulans. To identify the fkh gene(s) associated with fungal development, we examined mRNA levels of these six genes and found that the level of fkhB and fkhD mRNA was significantly elevated during asexual development and in conidia. To investigate the roles of FkhB and FkhD, we generated fkhB and fkhD deletion mutants and complemented strains and investigated their phenotypes. The deletion of fkhB, but not fkhD, affected fungal growth and both sexual and asexual development. The fkhB deletion mutant exhibited decreased colony size with distinctly pigmented (reddish) asexual spores and a significantly lower number of conidia compared with these features in the wild type (WT), although the level of sterigmatocystin was unaffected by the absence of fkhB. Furthermore, the fkhB deletion mutant produced sexual fruiting bodies (cleistothecia) smaller than those of WT, implying that the fkhB gene is involved in both asexual and sexual development. In addition, fkhB deletion reduced fungal tolerance to heat stress and decreased trehalose accumulation in conidia. Overall, these results suggest that fkhB plays a key role in proper fungal growth, development, and conidial stress tolerance in A. nidulans.

• 한국균학회소식:학술대회논문집
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• 한국균학회 2014년도 춘계학술대회 및 임시총회
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• pp.39-39
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• 2014

In a homothallic filamentous fungus Aspergillus nidulans, sexual and asexual developments are largely affected by the genetic and environmental factors. To regulate the complex subsets of genes involved in the developmental processes accurately, tight regulations of transcription factors are required. The forkhead type transcription factors are the class of regulators that function in a broad spectrum of cellular and developmental processes in many species from yeast to human. Here, we identified the fkhA and fkhB genes that encode a conserved forkhead transcription factors. The fkhA deletion resulted in the complete loss of fruiting body formation under all conditions favoring sexual development, suggesting that the fkhA gene is required for sexual development in A. nidulans. Overexpression of fkhA resulted in enhanced formation of fruiting bodies under induction condition not only in the normal condition but also in the condition of presence of 0.6 M KCl, which strongly inhibits sexual development. To know the function of the fkhB gene, we also generated fkhB knock-out strain in A. nidulans. Deletion of fkhB resulted in abnormal conidiophore formation under standard conditions and delayed sexual development process, suggesting that the fkhB gene plays an important role in conidiophore morphogenesis Taken together, these results suggest that the fkhA gene is necessary and sufficient for regulating sexual development and the fkhB gene is a transcription factor related in asexual developmental process in A. nidulans.

• Journal of Oral Medicine and Pain
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• 제48권2호
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• pp.39-44
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• 2023

Primary Sjögren's syndrome (pSS) is an autoimmune progressive disease characterized by dysfunction and inflammation of the salivary glands. The underlying mechanisms of salivary gland involvement in pSS remain unclear, and researchers have primarily focused on immunological phenomena, making it difficult to distinguish between the cause and effect of the disease. Consequently, our research aims to directly investigate changes in homeostasis occurring in acinar cells, specifically in the context of muscarinic signaling, mucins, aquaporins, and forkhead box protein O1, to elucidate the initial step of pSS. We compare the disease-related phenomena observed in salivary gland acinar cells in pSS with the overall process of salivary secretion.

• Asian-Australasian Journal of Animal Sciences
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• 제21권4호
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• pp.499-509
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• 2008

Foxo1 plays an important role in the integration of hormone-activated signaling pathways with the complex transcriptional cascade that promotes preadipocyte differentiation of clonal cell lines from rodents. We isolated the full-length cDNA of porcine FoxO1 gene using RACE, confirmed by visual Northern blotting. The deduced amino acids indicated 94% and 90% identities with the corresponding human and mice aa. Analysis of the aa sequence, showed that it included a Forkhead domain (aa 167-247), a transmembrane structure domain (aa 90-113), a LXXLL motif (aa 469-473), and 51 Ser, 8 Thr, and 4 Tyr phosphorylation sites, indicating a potential important role for FoxO1 transcriptional activity in vivo. Using the IMpRH panel, we mapped FoxO1 gene to chromosome 11p13. Our data provide basic molecular information useful for the further investigation on the function of FoxO1 gene. Time-course analysis of FoxO1 expressions indicated that levels of mRNA and protein gradually increased from day 0 to 3, and it reached almost maximal level at day 3, then decreased from day 5 to 7 in porcine primary preadipocyte differentiation. After induction by IGF-1, GPDH activity and accumulation of lipid increased, however, expressions of FoxO1 mRNA and protein were inhibited in a dose dependent manner. These results suggest that FoxO1 takes part in porcine preadipocyte differentiation and expressions of FoxO1 were regulated by IGF-1.

• Asian Pacific Journal of Cancer Prevention
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• 제15권24호
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• pp.10621-10625
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• 2015

Background: Increasing evidence has indicated that high Forkhead box protein C2 (FOXC2) level is closely associated with the development, progression, and poor prognosis of a variety of tumors. However, the relationship between FOXC2 and the progression of human gliomas remains to be clarified. The aim of present study was to assess FOXC2 expression and to explore its contribution in human gliomas. Materials and Methods: Realtime quantitative PCR was performed to examine FOXC2 expression in 85 pairs of fresh frozen glioma tissues and corresponding non-neoplastic brain tissues. Associations of FOXC2 expression with clinicopathological factors and prognosis of glioma patients were statistically analyzed. Results: The relative mRNA expression of FOXC2 was significantly higher in glioma tissues than the corresponding non-neoplastic brain tissues (p<0.001). In addition, high FOXC2 expression was significantly associated with advanced pathological grade (P=0.005) and the low Karnofsky performance score (KPS) (p=0.003), correlating with poor survival (p<0.001). Furthermore, multivariate Cox regression analysis showed that high FOXC2 expression was an independent predictor of overall survival (p=0.006). Conclusions: FOXC2 may act as an oncogenic gene and represent a potential regulator of aggressive development and a candidate prognostic marker in human gliomas.

• The Plant Pathology Journal
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• 제30권2호
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• pp.136-150
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• 2014

Although multiple transcription factors (TFs) have been characterized via mutagenesis to understand their roles in controlling pathogenicity and infection-related development in Magnaporthe oryzae, the causal agent of rice blast, if and how forkhead-box (FOX) TFs contribute to these processes remain to be characterized. Four putative FOX TF genes were identified in the genome of M. oryzae, and phylogenetic analysis suggested that two of them (MoFKH1 and MoHCM1) correspond to Ascomycota-specific members of the FOX TF family while the others (MoFOX1 and MoFOX2) are Pezizomycotina-specific members. Deletion of MoFKH1 (${\Delta}Mofkh1$) resulted in reduced mycelial growth and conidial germination, abnormal septation and stress response, and reduced virulence. Similarly, ${\Delta}Mohcm1$ exhibited reduced mycelial growth and conidial germination. Conidia of ${\Delta}Mofkh1$ and ${\Delta}Mohcm1$ were more sensitive to one or both of the cell cycle inhibitors hydroxyurea and benomyl, suggesting their role in cell cycle control. On the other hand, loss of MoFOX1 (${\Delta}Mofox1$) did not show any noticeable changes in development, pathogenicity, and stress response. Deletion of MoFOX2 was not successful even after repeated attempts. Taken together, these results suggested that MoFKH1 and MoHCM1 are important in fungal development and that MoFKH1 is further implicated in pathogenicity and stress response in M. oryzae.

• The Korean Journal of Physiology and Pharmacology
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• 제27권2호
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• pp.167-176
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• 2023

This study aims to explore the impact of Rehmannioside D (RD) on ovarian functions of rats with diminished ovarian reserve (DOR) and its underlying mechanisms of action. A single injection of cyclophosphamide was performed to establish a DOR rat model, and fourteen days after the injection, the rats were intragastrically administrated with RD for two weeks. Rat estrus cycles were tested using vaginal smears. Ovarian tissues were histologically evaluated, the number of primordial, mature, and atretic follicles was calculated, and the apoptotic rate of granulosa cells. Follicle-stimulating hormone (FSH), luteinizing hormone (LH), and estradiol (E₂) levels were determined by ELISA assays. Protein levels of Forkhead Box O1 (FOXO1), KLOTHO, Bcl-2, and Bax were investigated in ovarian tissues of DOR rats. The binding between FOXO1 and KLOTHO was verified by ChIP assay. High-dose administration of RD into DOR rats improved their estrus cycles, increased ovarian index, enhanced the number of primordial and mature follicles, reduced the number of atretic follicle number, and ovarian granulosa cell apoptosis in addition to inhibiting FSH and LH levels and upregulating E₂ expression. FOXO1 and KLOTHO were significantly suppressed in DOR rats. FOXO1 knockdown partially suppressed the protective effects of RD on DOR rats, and KLOTHO overexpression could restore RD-induced blockade of DOR development despite knocking down FOXO1. FOXO1 antibody enriched KLOTHO promoter, and the binding between them was reduced in DOR group compared to that in sham group. RD improved ovarian functions in DOR rats and diminished granulosa cell apoptosis via the FOXO1/KLOTHO axis.

• 대한의생명과학회지
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• 제24권1호
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• pp.15-22
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• 2018

Pristimerin is a triterpene compound isolated from plant extracts that reportedly possesses antitumor, antioxidant, and anti-inflammatory activities. The current study was designed to evaluate the antitumor effects of pristimerin on human colon cancer cells. Treatment of the human colon cancer cells, HCT116 and SW480, with pristimerin led to a dose-dependent decrease in cell proliferation. Flow cytometry experiments showed that pristimerin increased cell apoptotic rate and decreased the mitochondrial membrane potential in HCT116 and SW480 cells. Western blot assay showed that pristimerin induced increased cleavage of caspase-3, -7, -8, and poly ADP ribose polymerase. Treatment with pristimerin also caused a marked decrease in the expression of Bcl-2 and Bcl-xL. Additionally, the levels of phosphorylated AKT and forkhead box O3a (FOXO3a) were decreased in pristimerin-treated colon cancer cells. Taken together, our study illustrated that pristimerin promoted apoptosis via the AKT/FOXO3a signaling pathway in colon cancer cells, elucidating that it might be considered as a potential agent for colon cancer therapy.