Two ZNF509 (ZBTB49) isoforms induce cell-cycle arrest by activating transcription of p21/CDKN1A and RB upon exposure to genotoxic stress

ZNF509 is unique among POK family proteins in that four isoforms are generated by alternative splicing. Short ZNF509 (ZNF509S1, -S2 and -S3) isoforms contain one or two out of the seven zinc-fingers contained in long ZNF509 (ZNF509L). Here, we investigated the functions of ZNF509 isoforms in response to DNA damage, showing isoforms to be induced by p53. Intriguingly, to inhibit proliferation of HCT116 and HEK293 cells, we found that ZNF509L activates p21/CDKN1A transcription, while ZNF509S1 induces RB. ZNF509L binds to the p21/CDKN1A promoter either alone or by interacting with MIZ-1 to recruit the co-activator p300 to activate p21/CDKN1A transcription. In contrast, ZNF509S1 binds to the distal RB promoter to interact and interfere with the MIZF repressor, resulting in derepression and transcription of RB. Immunohistochemical analysis revealed that ZNF509 is highly expressed in normal epithelial cells, but was completely repressed in tumor tissues of the colon, lung and skin, indicating a possible role as a tumor suppressorope

Kr-pok increases FASN expression by modulating the DNA binding of SREBP-1c and Sp1 at the proximal promoter

Kr-pok (kidney cancer-related POZ domain and Krüppel-like protein) is a new proto-oncogenic POZ-domain transcription factor. Fatty acid synthase gene (FASN) encodes one of the key enzymes in fatty acids synthesis and is the only enzyme that synthesizes fatty acids in cancer cells. Sp1 and SREBP-1c are the two major transcription activators of FASN. We investigated whether Kr-pok modulates transcription of the FASN. FASN expression is significantly decreased in Kr-pok knockout murine embryonic fibroblasts. Coimmunoprecipitation, GST fusion protein pull-down, and immunocytochemistry assays show that the zinc-finger domain of Kr-pok interacts directly with the bZIP DNA binding domain of SREBP-1. Electrophoretic mobility shift assay, oligonucleotide pull-down, and chromatin immunoprecipitation assays showed that Kr-pok changes the transcription factor binding dynamics of Sp1 and SREBP-1c to the SRE/E-box elements of the proximal promoter. We found that Kr-pok expression increased during 3T3-L1 preadipocyte differentiation and that FASN expression is decreased by the knockdown of Kr-pok. Kr-pok facilitates the SREBP-1c-mediated preadipocyte differentiation and/or fatty acid synthesis. Kr-pok may act as an important regulator of fatty acid synthesis and may induce rapid cancer cell proliferation by increasing palmitate synthesis.ope

KR-POK interacts with p53 and represses its ability to activate transcription of p21WAF1/CDKN1A.

Transcriptional regulation by p53 is thought to play a role in its ability to suppress tumorigenesis. However, there remain gaps in understanding about how p53 regulates transcription and how disrupting this function may promote cancer. Here we report a role in these processes for the kidney cancer-related gene KR-POK (ZBTB7C), a POZ domain and Krüppel-like zinc finger transcription factor that we found to physically interact with p53. Murine embryonic fibroblasts isolated from genetically deficient mice (Kr-pok(-/-) MEFs) exhibited a proliferative defect relative to wild-type mouse embryonic fibroblasts (MEF). The zinc finger domain of Kr-pok interacted directly with the DNA binding and oligomerization domains of p53. This interaction was essential for Kr-pok to bind the distal promoter region of the CDKN1A gene, an important p53 target gene encoding the cell-cycle regulator p21WAF1, and to inhibit p53-mediated transcriptional activation of CDKN1A. Kr-pok also interacted with the transcriptional corepressors NCoR and BCoR, acting to repress histone H3 and H4 deacetylation at the proximal promoter region of the CDKN1A gene. Importantly, Kr-pok(-/-) MEFs displayed an enhancement in CDKN1A transactivation by p53 during the DNA damage response, without any parallel changes in transcription of either the p53 or Kr-pok genes themselves. Furthermore, Kr-pok promoted cell proliferation in vitro and in vivo, and its expression was increased in more than 50% of the malignant human kidney cancer cases analyzed. Together, our findings define KR-POK as a transcriptional repressor with a pro-oncogenic role that relies upon binding to p53 and inhibition of its transactivation function.ope

The proto-oncoprotein FBI-1 interacts with MBD3 to recruit the Mi-2/NuRD-HDAC complex and BCoR and to silence p21WAF/CDKN1A by DNA methylation

The tumour-suppressor gene CDKN1A (encoding p21Waf/Cip1) is thought to be epigenetically repressed in cancer cells. FBI-1 (ZBTB7A) is a proto-oncogenic transcription factor repressing the alternative reading frame and p21WAF/CDKN1A genes of the p53 pathway. FBI-1 interacts directly with MBD3 (methyl-CpG–binding domain protein 3) in the nucleus. We demonstrated that FBI-1 binds both non-methylated and methylated DNA and that MBD3 is recruited to the CDKN1A promoter through its interaction with FBI-1, where it enhances transcriptional repression by FBI-1. FBI-1 also interacts with the co-repressors nuclear receptor corepressor (NCoR), silencing mediator for retinoid and thyroid receptors (SMRT) and BCL-6 corepressor (BCoR) to repress transcription. MBD3 regulates a molecular interaction between the co-repressor and FBI-1. MBD3 decreases the interaction between FBI-1 and NCoR/SMRT but increases the interaction between FBI-1 and BCoR. Because MBD3 is a subunit of the Mi-2 autoantigen (Mi-2)/nucleosome remodelling and histone deacetylase (NuRD)-HDAC complex, FBI-1 recruits the Mi-2/NuRD-HDAC complex via MBD3. BCoR interacts with the Mi-2/NuRD-HDAC complex, DNMTs and HP1. MBD3 and BCoR play a significant role in the recruitment of the Mi-2/NuRD-HDAC complex– and the NuRD complex–associated proteins, DNMTs and HP. By recruiting DNMTs and HP1, Mi-2/NuRD-HDAC complex appears to play key roles in epigenetic repression of CDKN1A by DNA methylation.ope

ZBTB2 increases PDK4 expression by transcriptional repression of RelA/p65

The NF-κB is found in almost all animal cell types and is involved in a myriad of cellular responses. Aberrant expression of NF-κB has been linked to cancer, inflammatory diseases and improper development. Little is known about transcriptional regulation of the NF-κB family member gene RelA/p65. Sp1 plays a key role in the expression of the RelA/p65 gene. ZBTB2 represses transcription of the gene by inhibiting Sp1 binding to a Sp1-binding GC-box in the RelA/p65 proximal promoter (bp, -31 to -21). Moreover, recent studies revealed that RelA/p65 directly binds to the peroxisome proliferator-activated receptor-γ coactivator1α (PGC1α) to decrease transcriptional activation of the PGC1α target gene PDK4, whose gene product inhibits pyruvate dehydrogenase (PDH), a key regulator of TCA cycle flux. Accordingly, we observed that RelA/p65 repression by ZBTB2 indirectly results in increased PDK4 expression, which inhibits PDH. Consequently, in cells with ectopic ZBTB2, the concentrations of pyruvate and lactate were higher than those in normal cells, indicating changes in glucose metabolism flux favoring glycolysis over the TCA cycle. Knockdown of ZBTB2 in mouse xenografts decreased tumor growth. ZBTB2 may increase cell proliferation by reprogramming glucose metabolic pathways to favor glycolysis by upregulating PDK4 expression via repression of RelA/p65 expression.ope

확장된 계획행동이론을 통한 국민체력 100 참여자의 재참여 의도에 관한 연구

학위논문 (석사) -- 서울대학교 대학원 : 사범대학 체육교육과,글로벌스포츠매니지먼트전공, 2020. 8. Lee, Chung Gun.The average life expectancy of Koreans is increasing due to the development of medical science, improvements in life quality, and a general interest in health (KOSIS, 2018). However, although life expectancy has increased, the concept of health-adjusted life expectation (HALE) should also be considered. This is an indicator that estimates the average years a person can expect to live healthily. Some researchers think that HALE represents the best measurement for estimating the overall level of health for citizens. Consequently, the WHO (World Health Organization) use the concept as an official judgement criterion for yearly reports to provide information about the average level of healthy life for membership states (Lee et al., 2016). To live healthily and maintain quality of life, it is essential for people to manage physical fitness and participate in exercise. The effects of exercise have been proven by many research studies and the effects of exercise are apparent at all ages. The Korean government and the Ministry of Culture, Sports, and Tourism started the National Fitness Award (NFA; so-called Public Health 100 in Korea) program to promote sports and extend HALE by providing scientific measurement, exercise prescription, and eight weeks of exercise classes. After measurement, the Korea Sports Promotion Foundation (KSPO) give awards depending on the results. As well as Korea, other developed countries have designed sport programs based on life cycle and ages (Kim, 2014). The KSPO has operated the program since 2011, when an initial pilot program began with 4 centers and approximately 4,500 participants. Up to 2018, the KSPO and local government expanded the number of branches from 4 to 43, and the number of participants has increased to 267,729. The KSPO aims to add 8 more centers and increase the number of participants to approximately 300,000 (KSPO, 2019). However, according to the KSPO, only 0.56% of the population participated this program and one third of people who took part revisited the measurement. The NFA needs to expand further to promote healthy living and provide motivation to encourage participation in sport. The Korean government already spends a significant proportion of the budget (more than 10.7 billion won) on this program, which represents sports-related policy and the KSPO. This study was designed to identify factors that affect peoples intention to visit the scheme by applying the theory of planned behavior (TPB). This is one of the most well-known theories related to human behavior, and is used to predict exercise behavior, as well as for persuasive communication research, to structuralize attitudes, subjective norms, perceived control, intention, and behavior (Cho, 2012; Kim, Lee & Kim, 2003). However, some researchers indicate that the three components of TPB lack descriptive power for predicting human behavior (Armitage & Conner, 2001; Rivis & Sheeran, 2003). The need for new explanatory variables to mitigate the limitations of TPB led to the advanced and extended theory of planned behavior to include additional variables (Hagger et al., 2002; Kim, 2017). In this study, the concepts of spatial and temporal distance - aspects of the construal level theory (CLT) that affect intention and attitudes - are used as additional variables to extend TPB so that decision making in relation to revisiting the NFA scheme can be examined.본 연구의 목적은 확장된 계획행동이론(Extended Theory of Planned Behavior)을 활용하여 국민체력100(National Fitness Award) 참여자들의 재참여 의도를 살펴보는데 있다. 계획된 행동이론(Theory of Planned Behavior)의 태도, 사회적규범, 인지된 행동통제와 해석수준이론(Construal Level Theory)의 시간적, 공간적 거리가 재참여에 대한 태도 및 의도에 미치는 영향을 규명하였다. 해석수준이론의 시간적 거리는 미래의 상황이 현재로부터 시간적으로 얼마나 떨어져 있는지에 대한 주관적 인식이고, 공간적 거리는 미래의 상황이 나로부터 공간적으로 얼마나 멀리 떨어져 있는지에 대한 주관적 인식이다. 올림픽공원에 위치한 국민체력센터를 방문하는 방문객을 대상으로 설문을 실시하였으며, 수집된 총 226부의 설문지를 SPSS 22와 AMOS 22를 통해 확인적 요인분석 및 구조방정식 모형을 통해 분석을 실시하였다. 연구결과 계획된 행동이론의 구성요소 중 태도, 인지된 행동통제는 재참여 의도에 정(+)의 영향을 미치는 것으로 나타났다. 하지만 사회적 규범은 재방문 의도에 영향을 미치지 않는 것으로 나타났다. 시간적 거리의 경우, 재참여에 대한 태도와 의도에 정(+)의 영향을 미치는 것으로 나타났다. 즉 시간적으로 재측정까지 많이 남아있다고 느낄수록 가치 중심적인 상위해석수준이 작용하여 재참여에 대한 태도와 방문의도가 증가한다. 한편, 공간적 거리의 경우 재참여에 대한 태도 및 의도와 부적(-) 상관관계가 있는 것으로 나타났다. 참여자들이 국민체력센터가 가깝다고 느낄수록 구체성과 실행가능성 중심의 하위해석수준이 작용하여 재참여에 대한 태도와 의도가 증가하는 것이다. 이처럼 참여자가 지각하는 심리적 거리감에 따라 각기 다른 해석수준이 작용하게 되고, 해석수준을 토대로 커뮤니케이션 메세지를 전달하면 설득력을 높일 수 있다. 따라서 본 연구결과는 국민체력100 참가자들의 재참여 의사결정을 유도하고 프로그램 활성화를 위한 마케팅 전략을 수립하는데 도움을 줄 것으로 기대한다.Chapter 1. Introduction 1 1.1. Background and Necessity of the Research 1 1.2. Purpose of Research 7 1.3. Research hypothesis 7 Chapter 2. Theoretical Background 10 2.1. The Theory of Reasoned Action 11 2.2. The Theory of Planned Behavior 15 2.3. The extended Theory of Planned Behavior 19 2.4. Construal Level Theory and Psychological Distance 21 2.4.1. Temporal Distance 23 2.4.2. Spatial Distance 24 2.5. National Fitness Award Program Overview 25 2.5.1. Necessity and Effect of National Fitness Award 27 2.5.2. National Fitness Award Management System 29 2.5.3. National Fitness Award Program Measurement Grade 30 Chapter 3. Methodology 35 3.1. Sampling 35 3.2. Survey Instrument 38 3.3. Procedure 41 3.4. Statistical analysis 41 3.4.1. Reliability and discriminant validity analysis 41 3.4.2. Confirmatory factor analysis 42 3.4.3. Structure Equation Model 43 Chapter 4. Results 44 4.1. Descriptive analysis 44 4.2. Reliability and discriminant validity analysis 46 4.3. Confirmatory factor analysis 47 4.4. Hypothesis testing 48 Chapter 5. Discussion 52 5.1. Summary of key findings 52 5.2. Theoretical and practical implication 54 5.3. Limitations and future research directions 56 Bibliography 58 Appendix A 72 Appendix B 76 국문초록 80Maste

실험적 폐손상에서 유발가능 산화질소생산효소(iNOS)의 역할

Thesis(doctor`s)--서울대학교 대학원 :의학과 병리학전공,2007.Docto

ePRO-MP를 이용한 멀티프로세서 소프트웨어의 에너지와 성능 최적화

Thesis(masters) --서울대학교 대학원 :전기. 컴퓨터공학부, 2009.2.Maste

발암 단백질 PLZF와 PLZF-RARα의 분자수준에서의 기능 동정 및 작용점 규명

Dept. of Medical Science/박사Aberrant transcription repression through chromatin remodeling and histone deacetylation has been postulated as the driving force for tumorigenesis. PLZF (promyelocytic leukemia zinc finger) is initially isolated in fusion protein form with RARα (retinoic acid receptor alpha), which causes in APL (acute promyelocytic leukemia)-type leukemia. IHC(immunohistochemistry) of tumor microtissues arrays and oncomine data indicate that PLZF is aberrantly overexpressed in some human solid tumors, such as kidney renal carcinoma, testis seminoma, brain glioblastoma, and prostate adenocarcinoma. The data suggest that PLZF may be an oncoprotein involved in oncogenesis and cell proliferation. PLZF-RARα is considered as oncoprotein generated by chromosomal translocation between PLZF andRARα genes in the APL-type leukemic patients. Although PLZF-RARα was shown to cause leukemia and the molecular interaction between PLZF-RARα and corepressor-HDAC (Histon deacetylase) complex is important in oncogenesis, the target genes and action mechanism of PLZF-RARα remained largely unknown.I found that PLZF can repress transcription of the p21WAF/CDKN1A gene encoding p21, by binding to the proximal Sp1-binding GC-box 5/6 and the distal p53-responsive elements of the CDKN1A promoter. PLZF can also repress transcription of TP53 and destabilize p53 by deacetylation and ubiquitination, which also decreases transcription of CDKN1A. PLZF interacts with co-repressors, such as mSin3A, NCoR and SMRT, and deacetylateshistones H3 and H4 of the nucleosomes around the p21WAF/CDKN1A promoter. PLZF is an oncoprotein that causes cellular transformation and promotes cell proliferation in the HEK293A, HCT116, and H460 cells. PLZF does not induce apoptosis. PLZF-RARα promotes cell proliferation by repressing the transcription of the CDKN1A gene. Molecular characterization of oncoprotein PLZF-RARα revealed molecular mechanism similar to thePLZF, but PLZF-RARα also showed unique features in transcriptional repression PLZF-RARα is a competitive transcription repressor of p53, RARα and Sp1 and was shown to bind the proximal Sp1-binding GC-boxes 3, 4, 5/6, the RARE and the distal p53-responsive elements of CDKN1A promoter. PLZF-RARα also interacted with co-repressors, such as mSin3A, NCoR and SMRT, thereby deacetylates histones H3 and H4 of the nucleosomes around the CDKN1A promoter. Additionally, I found that PLZFRARα interacts with MBD3-NuRD complex, which leads to epigenetic silencing of the CDKN1A by DNA methylation and heterochromatinformation. PLZF and PLZF-RARα are the oncoproteins that repress transcription of the CDKN1A of the p53 pathway, by direct binding competition to the key regulatory elements such as proximal GC-boxes, distal p53 binding elements and RARE with Sp1, p53, and RARα. PLZF and PLZFRARα also represses transcription of CDKN1A by repressing transcription ofTP53 and by controlling the stability of p53.prohibitio

새로운 proto-oncogene인 FBI-1(pokemon/ZBTB7a)는 cyclin-d

Dept. of Medical Science/석사[한글]FBI-1 (Pokemon/ZBTB7a)는 여러 조직에서 발현되는 POK 계열의 전 발암 유전자이다. 우리는 FBI-1의 생체 내 기능을 연구하는 과정에서, 세포 주기 조절에서 중요한 조절 경로인 Arf-Mdm2-p53-p21 (p53 경로라 부름) 의 모든 유전자가 FBI-1에 의하여 그 발현이 억제되는 표적임을 발견하였다. FBI-1이 수행하는 세포 주기 조절 기전을 연구하고자, p53 경로의 Arf와 p53의 하부 유전자로써 세포 주기 조절의 핵심 존절자인 p21Waf/Cip1을 선택하여 연구를 진행하였다. 우리는 FBI-1, p53, Sp1, FRE(FBI-1 결합 부위)/Sp1-3 GC-box, 그리고 p53 결합 부위를 포함하는 복잡한 분자적 기전에 의하여 p21 유전자의 발현을 FBI-1이 억제 조절함을 발견하였다. 구체적으로 p21 프로모터에서 FBI-1은 Sp1에 의한 전사 활성화에 중요하다고 보고된 Sp1-3 GC-box (FBI-1 결합 부위와 중첩됨) 에서 전사 활성자인 Sp1과 분자적인 자리 경쟁에 의해 전사를 억제하며, 이를 통해서 Sp1과 p53간의 상호작용에 의한 전사 활성화도 억제한다. 또한 FBI-1이 p21 유전자의 전사 억제를 위해 distal p53 결합 부위에 결합하여 전사 활성화시키는 p53과 경쟁적으로 결합하는 것을 또한 발견하였다. FBI-1은 일단 proximal, distal 조절 부위에 결합하면, 전사 억제에서 중요한 HDAC이 포함된 mSIN3A, N-CoR, SMRT과 같은 보조전사 억제인자들과 상호작용하여, p21 유전자의 proximal 프로모터 부위에서 acetylation된 histon H3과 histon H4를 deacetylation함으로 nucleosome의 구조를 촘촘하게 하여 전사를 억제를 한다.나아가 우리는 p53 경로, 특히 p21의 전사 억제가 의미하는 생물학적기능을 연구하고자 세포주기의 FACS 분석, Foci-formation 분석, BrdU-incorporation 속도 등을 조사한 결과, FACS 분석결과, DNA 합성기 (S-상) 세포의 수가 현저히 증가함을 보였고, 이는 BrdU incorporation 속도와 일치하여 FBI-1은 증식이 빠른 세포로의 형질 전환을 유도하고 세포 주기는 촉진시킴을 발견하였다. 여러 병리 암 조직의 면역 화학 염색에서는 FBI-1이 adenocarcinoma와 squamous cell carconoma에서 두드러지게 발현되는 것을 관찰되었다.결론적으로 FBI-1은 Arf-Mdm2-p53-p21 (p53 경로) 경로의 중요한 억제조절 전사 인자이며 이를 통해 세포를 암세포로 형질전환 시키며 세포의 증식을 촉진함을 발견하였다. 우리의 연구는 어떻게 새로운 전 발암 유전자인 FBI-1이 암세포로의 전환을 유발하고, 종양세포의 증식을 유도하는지를 설명해주고 있다. [영문]FBI-1 (Pokemon/ZBTB7a) is a ubiquitous POK family oncogenic transcription factor. Virtually every gene in the Arf-Mdm2-p53-p21 regulatory pathway, important in the regulation of cell cycle progression, was found to be a repression target of FBI-1. In this study, the cell cycle regulator gene p21Waf/Cip1, which is downstream of Arf and p53, was selected for investigation into the molecular mechanism of transcriptional repression. This repression is known to involve FBI-1, p53, Sp1, the FRE (FBI-1 binding site)/Sp1-3 GC-box, and a distal p53-binding element. The FRE and Sp1-3 GC box sequences were found to overlap, and FBI-1 was found to represses transcription by molecular competition with Sp1 for binding to the Sp1-3 GC-box, an element critical in transcriptional regulation by Sp1 and synergistic transcriptional activation by Sp1 and p53. FBI-1 was also found to compete with p53 for binding to the distal p53-binding element to repress transcription. FBI-1 bound to sequences from both the proximal and distal regulatory elements. FBI-1 also interacted with corepressors such as mSin3A, N-CoR, and SMRT, and such interactions led to deacetylation of the Ac-H3 and Ac-H4 histones at the proximal promoter, which is likely an important step in transcriptional repression.FBI-1 expression caused cellular transformation and promoted cell cycle proliferation, as seen by a significant increase in the number of cells in the S-phase. Immunohistochemical staining showed that FBI-1 is prominently detected in adenocarcinoma (particularly in colon cancer) and squamous cell carcinoma cells, which express only low levels of p21. Overall, data from this study suggests that FBI-1 is the major regulator of the Arf-Mdm2-p53-p21Waf/Cip1 pathway and that FBI-1 promotes oncogenic transformation and tumor cell growth.ope