• KSBB Journal
• /
• 제24권3호
• /
• pp.217-226
• /
• 2009

p53은 전사인자로서 세포의 사멸이나 세포주기 조절 등 다양한 세포 활성을 보이기 때문에 일반적인 환경에서는 매우 낮은 수준으로 단백질 양이 확인된다. p53의 단백질 양과 활성은 다양한 세포 내 신호에 의하여 이루어지는 후전사 변형을 통하여 조절 받는다. 이중 유비퀴틴화는 세포 내에서 p53 단백질의 발현 수준이 낮게 유지되는 것이 가능하게 하는 대표적인 기전이다. 이러한 기전을 일으키는 대표적인 p53의 E3 ligase로는 mdm2, Pirh2, COP1, ARF-BP1 등이 보고되어 있으며, 각각 negative feedback loop나 다른 기전을 통하여 p53 단백질의 분해를 유도하여 세포의 항상성을 조절한다. 이 밖에도 p53은 mdm2나 WWP1, UBC13, MSL2와 같은 E3 ligase로 인해서 모노 유비퀴틴화 되고, p53의 세포 내 위치가 조절되어 전사인자로서의 활성이 억제된다. p53의 세포 내 위치와는 관계없이 p53의 전사인자로써의 활성 또한 아세틸화와 유비퀴틴화의 경쟁적 반응으로 인해 조절 될 수 있다. E4F1에 의한 유비퀴틴화는 세포주기와 관련된 유전자의 발현을 증가시키되 세포사멸 관련 유전자의 발현은 감소시키는 것으로 보아 p53의 수많은 downstream gene의 발현 또한 유비퀴틴화를 통해 조절 될 수 있음이 제시되었다. 앞으로의 연구는 신규 E3 ligase에 의한 p53의 유비퀴틴화 기전 연구 뿐 아니라 이와 관련된 다른 변형과의 관계에 대한 연구 또한 매우 중요하게 부각되어 질 것으로 예상된다.

• 한국작물학회:학술대회논문집
• /
• 한국작물학회 2017년도 9th Asian Crop Science Association conference
• /
• pp.149-149
• /
• 2017

The RING (Really Interesting New Gene) finger proteins are known to play crucial roles in various abiotic stresses in plants. In this study, we report on RING finger E3 ligase, ${\underline{O}ryza}$ ${\underline{s}ativa}$ ${\underline{s}alt$-, ${\underline{A}BA}$- and ${\underline{d}rounght}$ stress-${\underline{i}nduced}$ RING finger ${\underline{p}}rotein{\underline{1}}$ gene (OsSAD1). In vitro ubiquitination assay demonstrated that unlike OsSAD1, a single amino acid substitution ($OsSAD1^{C168A}$) of the RING domain showed no E3 ligase activity, supporting the notion that the activity of most E3s is specified by a RING domain. Result of Yeast-Two hybridization, In vivo protein degradation assay supports that OsSAD1 interacting with 3 substrate, OsSNAC2, OsGRAS44 and OsPIRIN1, and mediates proteolysis of 3 substrates via the 26S proteasome pathway. Subcellular localizations of OsSAD1 while approximately 62% of transient signals were detected in cytosol, 38% of signals were showed nucleus. However, transiently expression of OsSAD1 was detected in cytosol 30% while as 70% of nucleus under 200 mM salt treated rice protoplasts. Results of bimolecular fluorescence complementation (BiFC) showed that two nucleus-localized proteins (OsSNAC2 and OsGRAS44) interacted with OsSAD1 in the both cytosol and nucleus. Heterogeneous overexpression of OsSAD1 Heterogeneous overexpresssion of OsSAD1 in Arabidopsis exhibited sensitive phenotypes with respect to Salt-, mannitol-responsive seed germination, seedling growth. In ABA conditions, OsSAD1 overexpression plants showed highly tolerance phenotypes, such as root length and stomatal closure. Our findings suggest that the OsSAD1 may play a negative regulator in salt stress response by modulating levels of its target proteins.

• 한국발생생물학회지:발생과생식
• /
• 제16권3호
• /
• pp.169-175
• /
• 2012

Angelman syndrome (AS) is a neurodevelopmental disorder characterized by intellectual disability and autism. The genetic cause is the absence of UBE3A, an E3 ubiquitin ligase, from the maternal chromosome which can arise from multiple origins. Recently discovered targets of Ube3a are important for activity dependent changes in synaptic transmission and spine morphology. Plasticity studies in an AS mouse model is important for basic plasticity research with regard to understanding protein homeostasis as well as the search for therapeutic targets for the patients. The progress on synaptic plasticity from this unique disorder is reviewed.

• Animal cells and systems
• /
• 제16권4호
• /
• pp.280-288
• /
• 2012

Targeted degradation of proteins through ubiquitin-mediated proteolysis is an important control mechanism in various cellular processes. The process of ubiquitin conjugation is achieved by three enzyme complexes, among which the ubiquitin ligase complex (E3) is in charge of substrate specificity. The SCF (SKP1-CUL1-F-box) family portrays the largest and the most characterized member of the E3 ligases. For each SCF complex, the ubiquitination target is recognized by the F-box protein subunit, which interacts with the substrate through a unique C-terminal domain. We have characterized a novel F-box protein CFL-1 that represents a single LRR-type F-box (FBXL) in the Caenorhabditis elegans genome. CFL-1 is highly homologous to FBXL20 and FBXL2 of mammals, which are known to regulate synaptic vesicle release and cell cycle, respectively. A green fluorescence protein (GFP)-reporter gene fused to the cfl-1 promoter showed restricted expression around the amphid and the anus. Modulation of CFL-1 activity by RNAi affected the time interval between defecations. RNAi-treated worms also exhibited reduced tendency to form dauer when exposed to daumone. The potential involvement of CFL-1 in the control of defecation and pheromone response adds to the ever expanding list of cellular processes controlled by ubiquitin-mediated proteolysis in C. elegans. We suggest that CFL-1, as a single LRR-type F-box protein in C. elegans, may portray a prototype gene exerting diverse functions that are allocated among multiple FBXLs in higher organisms.

• Molecules and Cells
• /
• 제24권3호
• /
• pp.445-451
• /
• 2007

Translation initiation factor 4E (eIF4E) is a key regulator of protein synthesis. Abnormal regulation of eIF4E is closely linked to oncogenic transformation. Several regulatory mechanisms affecting eIF4E are discussed, including transcriptional regulation, phosphorylation and binding of an inhibitor protein. However it is not clear how the level of eIF4E protein is regulated under basal conditions. Here we demonstrate that Diap1 (Drosophila Inhibitor of Apoptosis Protein), a cell death inhibitor, binds directly to eIF4E and poly-ubiquitinates it via its E3 ligase activity, promoting its proteasome-dependent degradation. Expression of Diap1 caused a reduction of Cyclin D1 protein level and inhibited the growth stimulation induced by overexpression of eIF4E. Taken together, our results suggest that the level of eIF4E protein is regulated by Diap1, and that IAPs may play a role in cap-dependent translation by regulating the level of eIF4E protein.

• 한국발생생물학회지:발생과생식
• /
• 제7권2호
• /
• pp.69-80
• /
• 2003

대량의 발생 유전학적 연구가 가능한 척추동물로서 최근 제브라피쉬가 새로운 동물모델로 급부상하고 있다 다양한 형태의 돌연변이들로부터 새로운 유전자들이 발굴되어지고 있으며, 인간 유전체의 기능 분석 수단으로 활용되어지고 있다. 신경계의 형성과 분화에 이상이 있는 hendless와 mind bomb이라는 두 가지 돌연변이주에서 positional cloning에 의한 원인 유전자의 발굴과 기능 분석의 예로써 현재 제브라피쉬의 연구 현황을 살펴보고자 한다. headless의 원인 유전자로 Tcf-3가 밝혀졌으며, 초기 발생단계에서 Wnt 신호전달이 두뇌의 형태형성과 영역 결정에서 핵심적 역할을 하고 있다는 사실이 밝혀졌다. mind bomb에서의 비정상적인 신경세포의 운명 결정은 lateral inhibition과 Notch 신호전달의 결함에 의한 것이고, 그 원인 유전자는 Notch ligand인 Delta에 결합하는 새로운 ubiquitin E3 ligase로 밝혀졌다. 이러한 돌연변이를 통한 연구는 현재 인간 질환모델의 개발이라는 방향으로 확대되고 있다.

• Molecules and Cells
• /
• 제27권3호
• /
• pp.359-363
• /
• 2009

Chfr, a checkpoint with FHA and RING finger domains, plays an important role in cell cycle progression and tumor suppression. Chfr possesses the E3 ubiquitin ligase activity and stimulates the formation of polyubiquitin chains by Ub-conjugating enzymes, and induces the proteasome-dependent degradation of a number of cellular proteins, including Plk1 and Aurora A. While Chfr is a nuclear protein that functions within the cell nucleus, how Chfr is localized in the nucleus has not been clearly demonstrated. Here, we show that nuclear localization of Chfr is mediated by nuclear localization signal (NLS) sequences. To reveal the signal sequences responsible for nuclear localization, a short lysine-rich stretch (KKK) at amino acid residues 257-259 was replaced with alanine, which completely abolished nuclear localization. Moreover, we show that nuclear localization of Chfr is essential for its checkpoint function but not for its stability. Thus, our results suggest that NLS-mediated nuclear localization of Chfr leads to its accumulation within the nucleus, which may be important in the regulation of Chfr activation and Chfr-mediated cellular processes, including cell cycle progression and tumor suppression.

• 한국미생물·생명공학회지
• /
• 제7권3호
• /
• pp.165-173
• /
• 1979

자연선조사한 대장균B주의 세포분열회복활성분을 구명코저 자외선내성균인 대장균 B/r 주의 초음파추출액으로부터 활성성분分을 분리한 결과 $\beta$-NAD가 관여함이 발표되었다. 본고에서는$\beta$-NAD 이외 Magnesium이 활성물질의 안정화가 중요한 역할을 나타냄을 구명하였으며 10~30%의 서당밀 도구배원심분리에 의해 2 개의 새로운 활성부분이 있음을 확인하였다. 2 개의 활성물질 가운데 하나는 원심관의 최하부에 위치하였으며 또 다른 하나는 상부의 분자량 45,000 부위에서 회수되었다. 하부에 위치한 활성획분은 Mg++이 그 활성에 무관하였으나 상부의 저분자 활성부분은 $Mg^{++}$을 첨가하지 않으면 회수가 불가능하였다. 저분자활성부분은 pronase에 대해 감수성이었으며 DNA-ligase 는 아님이 추정되었다. 초원심분리과정에 $N_2$ gas를 처리할 경우 aeration에 비해 약 2 배의 활성이 나타났다. $Mg^{++}$은 $\beta$-NAD에 또하나의 회복활성 및 필수적인자로 요구된다고 생각된다.

• Molecules and Cells
• /
• 제41권3호
• /
• pp.168-178
• /
• 2018

Intracellular communication via ubiquitin (Ub) signaling impacts all aspects of cell biology and regulates pathways critical to human development and viability; therefore aberrations or defects in Ub signaling can contribute to the pathogenesis of human diseases. Ubiquitination consists of the addition of Ub to a substrate protein via coordinated action of E1-activating, E2-conjugating and E3-ligating enzymes. Approximately 40 E2s have been identified in humans, and most are thought to be involved in Ub transfer; although little information is available regarding the majority of them, emerging evidence has highlighted their importance to human health and disease. In this review, we focus on recent insights into the pathogenetic roles of E2s (particularly the ubiquitin-conjugating enzyme E2O [UBE2O]) in debilitating diseases and cancer, and discuss the tantalizing prospect that E2s may someday serve as potential therapeutic targets for human diseases.

• 한국동물학회:학술대회논문집
• /
• 한국동물학회 2001년도 한국생물과학협회 학술발표대회
• /
• pp.252.1-252
• /
• 2001

No Abstract, See Full Text