• BMB Reports
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• 제45권6호
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• pp.331-336
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• 2012

The retinoid-related orphan nuclear receptor gamma ($ROR{\gamma}$) plays critical roles in regulation of development, immunity and metabolism. As transcription factor usually forms a protein complex to function, thus capturing and dissecting of the $ROR{\gamma}$ protein complex will be helpful for exploring the mechanisms underlying those functions. After construction of the recombinant tandem affinity purification (TAP) plasmid, pMSCVpuro $ROR{\gamma}$-CTAP(SG), the nuclear localization of $ROR{\gamma}$-CTAP(SG) fusion protein was verified. Following isolation of $ROR{\gamma}$ protein complex by TAP strategy, seven candidate interacting proteins were identified. Finally, the heat shock protein 90 (HSP90) and receptor-interacting protein 140 (RIP140) were confirmed to interplay with $ROR{\gamma}$ by co-immunoprecipitation. Interference of HSP90 or/and RIP140 genes resulted in dramatically decreased expression of CYP2C8 gene, the $ROR{\gamma}$ target gene. Data from this study demonstrate that HSP90 and RIP140 proteins interact with $ROR{\gamma}$ protein in a complex format and function as co-activators in the $ROR{\gamma}$-mediated regulatory processes of HepG2 cells.

• Asian Pacific Journal of Cancer Prevention
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• 제14권10호
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• pp.5789-5795
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• 2013

Background: The high mobility group box 1 (HMGB1) protein is a widespread nuclear protein present in most cell types. It typically locates in the nucleus and functions as a nuclear cofactor in transcription regulation. However, HMGB1 can also localize in the cytoplasm and be released into extracellular matrix, where it plays critical roles in carcinogenesis and inflammation. However, it remains elusive whether HMGB1 is relocated to cytoplasm in clear cell renal cell carcinoma (ccRCC). Methods: Nuclear and cytoplasmic proteins were extracted by different protocols from 20 ccRCC samples and corresponding adjacent renal tissues. Western blotting and immunohistochemistry were used to identify the expression of HMGB1 in ccRCC. To elucidate the potential mechanism of HMGB1 cytoplasmic translocation, HMGB1 proteins were enriched by immunoprecipitation and analyzed by mass spectrometry (MS). Results: The HMGB1 protein was overexpressed and partially localized in cytoplasm in ccRCC samples (12/20, 60%, p<0.05). Immunohistochemistry results indicated that ccRCC of high nuclear grade possess more HMGB1 relocation than those with low grade (p<0.05). Methylation of HMGB1 at lysine 112 in ccRCC was detected by MS. Bioinformatics analysis showed that post-translational modification might affect the binding ability to DNA and mediate its translocation. Conclusion: Relocation of HMGB1 to cytoplasm was confirmed in ccRCC. Methylation of HMGB1 at lysine 112 might the redistribution of this cofactor protein.

• Journal of Ginseng Research
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• 제42권4호
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• pp.436-446
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• 2018

Background: The potential therapeutic values of Korean Red Ginseng extract (KRGE) in autoimmune disorders of nervous system have not been fully investigated. Methods: We used an acute experimental autoimmune encephalomyelitis animal model of multiple sclerosis and determined the effects and mechanism of KRGE on spinal myelination. Results: Pretreatment with KRGE (100 mg/kg, orally) for 10 days before immunization with myelin basic protein $(MBP)_{68-82}$ peptide exerted a protective effect against demyelination in the spinal cord, with inhibited recruitment and activation of immune cells including microglia, decreased mRNA expression of detrimental inflammatory mediators (interleukin-6, interferon-${\gamma}$, and cyclooxygenase-2), but increased mRNA expression of protective inflammatory mediators (insulin-like growth factor ${\beta}1$, transforming growth factor ${\beta}$, and vascular endothelial growth factor-1). These results were associated with significant downregulation of p38 mitogen-activated protein kinase and nuclear factor-${\kappa}B$ signaling pathways in microglia/macrophages, T cells, and astrocytes. Conclusion: Our findings suggest that KRGE alleviates spinal demyelination in acute experimental autoimmune encephalomyelitis through inhibiting the activation of the p38 mitogen-activated protein kinase/nuclear factor-${\kappa}B$ signaling pathway. Therefore, KRGE might be used as a new therapeutic for autoimmune disorders such as multiple sclerosis, although further investigation is needed.

• 생명과학회지
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• 제23권11호
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• pp.1311-1316
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• 2013

hnRNP L은 pre-mRNA에 결합하는 단백질들 중에서 핵심이 되는 단백질이다. hnRNP L은 양이 아주 많은 핵 단백질로서 핵과 세포질을 왕복하는 특성을 지니고 있다. 이 단백질은 염색질 변형(chromatin modification), pre-mRNA 스플라이싱, 인트론이 없는 유전자들에서 유래한 mRNA들의 세포질로의 반출(export), IRES-매개성 번역, mRNA의 안정성 조절, 정자형성과정 등, 세포 내의 여러 가지 과정에 관여하고 있는 것으로 알려져 있다. 이 논문에서는 hnRNP L과 결합하는 세포 내 단백질을 찾아내기 위하여 사람의 간세포 cDNA library를 사용하여 이스트 two-hybrid 탐색 실험을 수행하였다. 그 결과 사람의 간세포에서 IGFBP-4가 hnRNP L과 상호결합하는 새로운 파트너라는 것을 발견하였다. 본 연구를 통하여 hnRNP L이 이스트 two-hybrid 시스템에서 IGFBP-4와 특이적으로 상호 결합한다는 것을 처음으로 발견하였다. 본 연구에서는 또한 이스트 two-hybrid 시스템에서 hnRNP L이 IGFBP-4와 상호결합한다는 점을 in vitro pull-down 실험을 통하여 재확인하였다.

• International Journal of Industrial Entomology and Biomaterials
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• 제3권1호
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• pp.75-81
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• 2001

We have constructed a novel recombinant Bombyx mori nuclear polyhedrosis virus (BmNPV) producing the green fluorescent polyhedra. For the production of the fluorescent polyhedra, partial polyhedrin gene containing KRKK as nuclear localization site from the BmNPV polyhedrin gene and the green fluorescent protein (gfp) gene were introduced under the control of p10 promoter of BmNPV. The recombinant BmNPV was stably produced fluorescent polyhedra in the infected Bm5 cells and the morphology of the fluorescent polyhedra was similar to that of wild-type BmNPV. The fluorescent polyhedra had 32 kDa native polyhedrin and 41 kDa fusion protein. From these data, we have further developed a novel BmNPV p10-based transfer vector producing recombinant polyhedra with foreign gene Product. The novel BmNPV P10-based transfer vector is composed of partial polyhedrin gene, factor Xa, and multiple cloning sites.

• 한국응용곤충학회지
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• 제30권2호
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• pp.144-149
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• 1991

국내에서 분리된 파밤나방 핵다각체병바이러스(Spodoptera exigua nuclear polyhedrosis virus: SeNPV)의 생화학적인 특성을 규명하기 위하여 몇가지 실험을 행하였다. SeNPV는 하나의 envelopeso내에 다수의 nucleocapsid가 존재하는 MNPV(multiple embeded NPV)형태였다. 다각체단백질은 분자량 30kb의 단일 band로 나타났으며, Spodoptera litura NPV와 Bombyx mori NPV의 다각체단백질 항체에 반응하여 뚜렷한 침강선을 형성하였다. 비리온 단백질을 은염색한 결과, 많은 수의 minor band들이 포함된 49개의 band로 나타났으며, 바이러스 DNA를 분리하여 여러종의 제한효소에 의한 대략적인 genome size는 약 110kb 였다.

• Molecules and Cells
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• 제26권3호
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• pp.270-277
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• 2008

This study addresses the physiological functions of the Ran-binding protein homolog NbRanBP1 in Nicotiana benthamiana. Virus-induced gene silencing (VIGS) of NbRanBP1 caused stunted growth, leaf yellowing, and abnormal leaf morphology. The NbRanBP1 gene was constitutively expressed in diverse tissues and an NbRanBP1:GFP fusion protein was primarily localized to the nuclear rim and the cytosol. BiFC analysis revealed in vivo interaction between NbRanBP1 and NbRan1 in the nuclear envelope and the cytosol. Depletion of NbRanBP1 or NbRan1 reduced nuclear accumulation of a NbBTF3:GFP marker protein. In the later stages of development, NbRanBP1 VIGS plants showed stress responses such as reduced mitochondrial membrane potential, excessive production of reactive oxygen species, and induction of defense-related genes. The molecular role of RanBP1 in plants is discussed in comparison with RanBP1 function in yeast and mammals.

• 한국수정란이식학회지
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• 제30권1호
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• pp.33-43
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• 2015

Successful somatic cell nuclear transfer (SCNT) has been reported across a range of species using a range of recipient cells including enucleated metaphase II (MII) arrested oocytes, enucleated activated MII oocytes, and mitotic zygotes. However, the frequency of development to term varies significantly, not only between different cytoplast recipients but also within what is thought to be a homogenous population of cytoplasts. One of the major differences between cytoplasts is the activities of the cell cycle regulated protein kinases, maturation promoting factor (MPF) and mitogen activated protein kinase (MAPK). Dependent upon their activity, exposure of the donor nucleus to these kinases can have both positive and negative effects on subsequent development. Co-ordination of cell cycle stage of the donor nucleus with the activities of MPF and MAPK in the cytoplast is essential to avoid DNA damage and maintain correct ploidy. However, recent information suggests that these kinases may also effect reprogramming of the somatic nucleus and preimplantation embryo development by other mechanisms. This article will summarise the differences between cytoplast recipients, their effects on development and discuss the potential role/s of MPF and or MAPK in nuclear reprogramming.

• Animal cells and systems
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• 제12권3호
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• pp.143-148
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• 2008

Heterogeneous nuclear ribonucleoprotein E1(hnRNP E1) is one of the primary pre-mRNA binding proteins in human cells. It consists of 356 amino acid residues and harbors three hnRNP K homology(KH) domains that mediate RNA-binding. The hnRNP E1 protein was shown to play important roles in mRNA stabilization and translational control. In order to enhance our understanding of the cellular functions of hnRNP E1, we searched for interacting proteins through a yeast two-hybrid screening while using HeLa cDNA library as target. One of the cDNA clones was found to be human ribosomal protein L18a cDNA(GenBank accession number BC071920). We demonstrated in this study that human ribosomal protein L18a, a constituent of ribosomal protein large subunit, interacts specifically with hnRNP E1 in the yeast two-hybrid system. Such an interaction was observed for the first time in this study, and was also verified by biochemical assay.

• 한국잠사곤충학회지
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• 11호
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• pp.59-62
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• 1970

1. 한국에서 작잠농병을 발생시키는 주된 Virus는 핵질다색체 Virus 이다. 2. Virus 묶음은 봉입체단백직의 분자구조내에 함입되어있다. 3. 봉입체단백질에 존재하는 Virus묶음은 평균4개의 간상형 Virus입자로 되어있으며 이를 싸고있는 막은 두개로 되어있다. 4. 봉입체단백질내에 존재하는 Virus묶음은 질서와 균형있게 배열된 것이 아니고 되는데로 산재해 있는 것 같다. 5. Virus입자와 봉입예단백질은 전염된 세포의 염색체에서 형성된 소위 "Virogenic stroma"에서 생겨진것이다.