Phosphorylation of Ser246 residue in intergin-linked kinase 1 by serum- and glucocorticoid-induced kinase 1 is required to from a protein-protein complex with 14-3-3

Jaesun Chun,강상순 한국통합생물학회 2005 Animal cells and systems Vol.9 No.3

Integrin-linked kinase 1 (ILK1) regulates several protein kinases, including PKB/Akt kinase and glycogen synthase kinase . ILK1 is also involved distinctively in the cell morphological and structural functions by interacting with the components of the extracellular matrix or integrin. According to the information of serum- and glucocorticoid-induced kinase 1 (SGK1) substrate specificity (R-X-R-X-X-(S/T)-;  indicates a hydrophobic amino acid), two putative phosphorylation sites, Thr181 and Ser246, were found in ILK1. We showed that ILK1 fusion protein and two fluorescein-labeled ILK1 peptides, FITC-174RTRPRNGTLNl83 and FITC-239CPRLRIFSHP248, were phosphorylated by SGK1 in vitro. We also identified that 14-3-3 , and  among several 14-3-3 isotypes (and ) formed protein complex with ILK1 in COS-1 cells. Furthermore, the phosphorylation of Ser246 by SGK1 induced the binding with 14-3-3. It was also demonstrated that 14-3-3-bound ILK1 has reduced kinase activity. Thus, these data suggest that SGK1 phosphorylates Thr181 and Ser246 of ILK1 and the phosphorylation of its Ser246 makes ILK1 bind to 14-3-3, resulting in the inhibition of ILK1 kinase activity.

Ubiquitylation of Fe65 adaptor protein by neuronal precursor cell expressed developmentally down regulated 4-2 (Nedd4-2) via the WW domain interaction with Fe65

이은정,현성희,Jaesun Chun,신성화,강상순 생화학분자생물학회 2009 Experimental and molecular medicine Vol.41 No.8

Fe65 has been characterized as an adaptor protein, originally identified as an expressed sequence tag (EST) corresponding to an mRNA expressed at high levels in the rat brain. It contains one WW domain and two phosphotyrosine interaction/phosphotyrosine binding domains (PID1/PID2). As the neuronal precursor cell expressed developmentally down regulated 4-2 (Nedd4-2) has a putative WW domain binding motif (72PPLP75) in the N-terminal domain, we hypothesized that Fe65 associates with Nedd4-2 through a WW domain interaction, which has the characteristics of E3 ubiquitin-protein ligase. In this paper, we present evidence for the interaction between Fe65 WW domain and Nedd4-2 through its specific motif, using a pull down approach and co-immunoprecipitation. Additionally, the co-localization of Fe65 and Nedd4-2 were observed via confocal microscopy. Co-localization of Fe65 and Nedd4-2 was disrupted by either the mutation of Fe65 WW domain or its putative binding motif of Nedd4-2. When the ubiquitin assay was performed, the interaction of Nedd4-2 (wt) with Fe65 is required for the cell apoptosis and the ubiquitylation of Fe65. We also observed that the ubiquitylation of Fe65 (wt) was augmented depending on Nedd4-2 expression levels, whereas the Fe65 WW domain mutant (W243KP245K) or the Nedd4-2 AL mutant (72PPLP75 was changed to 72APLA75) was under-ubiquitinated significantly. Thus, our observations implicated that the protein-protein interaction between the WW domain of Fe65 and the putative binding motif of Nedd4-2 down-regulates Fe65 protein stability and subcellular localization through its ubiquitylation, to contribute cell apoptosis. Fe65 has been characterized as an adaptor protein, originally identified as an expressed sequence tag (EST) corresponding to an mRNA expressed at high levels in the rat brain. It contains one WW domain and two phosphotyrosine interaction/phosphotyrosine binding domains (PID1/PID2). As the neuronal precursor cell expressed developmentally down regulated 4-2 (Nedd4-2) has a putative WW domain binding motif (72PPLP75) in the N-terminal domain, we hypothesized that Fe65 associates with Nedd4-2 through a WW domain interaction, which has the characteristics of E3 ubiquitin-protein ligase. In this paper, we present evidence for the interaction between Fe65 WW domain and Nedd4-2 through its specific motif, using a pull down approach and co-immunoprecipitation. Additionally, the co-localization of Fe65 and Nedd4-2 were observed via confocal microscopy. Co-localization of Fe65 and Nedd4-2 was disrupted by either the mutation of Fe65 WW domain or its putative binding motif of Nedd4-2. When the ubiquitin assay was performed, the interaction of Nedd4-2 (wt) with Fe65 is required for the cell apoptosis and the ubiquitylation of Fe65. We also observed that the ubiquitylation of Fe65 (wt) was augmented depending on Nedd4-2 expression levels, whereas the Fe65 WW domain mutant (W243KP245K) or the Nedd4-2 AL mutant (72PPLP75 was changed to 72APLA75) was under-ubiquitinated significantly. Thus, our observations implicated that the protein-protein interaction between the WW domain of Fe65 and the putative binding motif of Nedd4-2 down-regulates Fe65 protein stability and subcellular localization through its ubiquitylation, to contribute cell apoptosis.

친밀감 정도 및 리더 유무에 따른 소집단 구성에서 나타나는 중학생들의 사회 관련 과학쟁점(SSI) 논변활동 양상 분석

(Su-Jin Lee),천재순(Jaesun Chun) 학습자중심교과교육학회 2017 학습자중심교과교육연구 Vol.17 No.24

본 연구는 소집단 구성 방식에 따른 중학생들의 사회 관련 과학쟁점(SSI) 논변활동 양상 분석에 그 목적이 있다. 연구 참여자는 S시에 위치한 E중학교의 과학반 학생 12명이다. 학교에서 많이 사용하는 소집단 구성 방식 중 세 가지를 선정하였다. 각각 친밀감 기반 구성, 무작위 구성, 친밀감에 기반한 리더 중심 소집단 구성이다. 논변활동의 주제는 모두 생명 영역으로 한정하였다. 한 학기 동안 총 5회의 논변활동을 실시하였다. 자료 수집은 소집단 논변활동 녹화 및 녹음을 통한 전사자료, 학생활동지, 교사의 관찰일지, 면담 자료 등을 통해 이루어졌다. 여러 자료들을 종합적으 로 검토하여 귀납적으로 분석하였으며, 삼각 측정법을 사용하여 연구의 신뢰도를 확보하였다. 소집단 구성 방식에 따른 논변활동 활성화 및 논변의 수준은 친밀감에 기반한 리더 중심 구성에서 가장 높았으며, 친밀감 기반 구성, 무작위 구성 순으로 높 게 나타났다. 소집단 구성 방식의 변화가 개인의 논변활동 활성화에 큰 차이를 나타내기도 하였다. 본 연구 결과는 소집단 구성 방식이 중학생들의 SSI 논변활동의 활성화 및 논변의 수준에 영향을 미침을 시사하고 있다. The purpose of this study is to analyze the aspect of the socio-scientific issue argumentation among middle school students based on methods of small group structure. The participants of this study were 12 students in science class at E middle school in S city. We selected three different methods for small group construction that are mostly used in schools: an intimacy-based small group, a randomized small group, and a leader-centered and intimacy-based small group. The subject of the argumentation were all limited to the area of life. Total five times of argumentation ware held throughout one semester. All discourse activities from each small group were videotaped and recorded for data collection and transcribed. Additional resources for analysis include student activity sheets, teacher observation logs, and interviews with all students immediately after argumentation. All the data were comprehensively analyzed and the reliability of the study was secured by using triangulation method of data. Our analysis showed that the leader-centered and intimacy-based small group was the most active in argumentative activities and the highest in the level of their argumentation as being analyzed in the perspective of resistance among three small groups. On the other hand, the randomized small group revealed the lowest level in both. The changes in the way of small group structure showed a big difference in activating the argumentative activity of the individual. The results of this study suggest that the methods of small group structure affect on both active participation in argumentation and the level of argumentation to middle school students.

질병 관련 직업을 주제로 한 진로탐색 프로그램 개발 및 적용: 중학생을 대상으로

조광희(Cho, Kwanghee),천재순(Chun, Jaesun) 학습자중심교과교육학회 2018 학습자중심교과교육연구 Vol.18 No.21

본 연구는 활동 중심의 질병 관련 진로탐색 프로그램을 개발하고, 프로그램 적용후 학생들의 질병 관련 진로탐색 경험을 현상학적으로 이해하는 데 목적을 두었다. 이를 위해 ADDIE 모형을 토대로 질병, 직업, 활동 요소로 구성한 프로그램을 개발 하고, 자유학기제를 시행하는 D광역시 B중학교에 적용하였다. 연구 참여자는 중학교 2학년 남학생 20명이며, 이들 중 7명의 학생들이 면담에 참여하였다. 학생들의 과학 관련 진로탐색 경험을 확인하기 위해 진로탐색 프로그램의 학생 수업참여 양상을 녹화 및 관찰 기록하였다. 과학교육전문가 2인으로부터 안면타당도를 받아 반구조화 면담을 진행하였으며, 진로탐색 프로그램 종료 후 만족도 조사를 실시하였다. 연구 결과 진로탐색 프로그램 개발 및 적용을 통해 학생들은 경험하지 못했던 다양한 직업들을 학교 내에서 실질적으로 경험하고 탐색할 수 있었다. 나아가 학생들은 체험 활동을 통해 자신의 진로탐색 경험을 재구성하였다. 본 프로그램의 만족도 조사 결과는 Likert 5점 척도로 4.86이다. The purpose of this study is to develop a activity-centered program for career exploration related diseases and to phenomenologically understand students’ experience of activities about career exploration related disease. The program based on the ADDIE model was developed to be composed of elements: diseases, careers, activities, and applied to free learning semester at B middle school in D city. The program participants were 20 students and 7 students of them were interviewed. Recording and describing were conducted to be confirmed the students’ experience of career exploration related diseases. Semi-structured interviews were conducted, after it had held with the validity from two science education experts. The students could experience and explore various jobs in careers which were not able to experience in the school through the career-exploration program. Furthermore, students have reconstricted their experience of the career exploration by participating in the activity for experience. The satisfaction survey of the program was 4.86 on the 5 point Likert scale.

농업계 특성화고 학생과 일반계고 학생의 과학의 본성에 대한 인식 및 과학 동기 수준의 비교분석

한영희(Younghee Han),천재순(Jaesun Chun) 학습자중심교과교육학회 2017 학습자중심교과교육연구 Vol.17 No.17

The level of agricultural specialized high school students in the nature of science and the science motivation was investigated and some of them were interviewed to get their specific thoughts about science and its class in this study . It is also conducted with the aim of looking for ways on how to support them in the learning of the aforementioned subject. For this purpose, the test for the nature of science, SUSSI, and the test for the science motivation, SMQⅡ, were conducted to the second year students of the schools which are located in the central region. They are 171 agricultural specialized high school students and 235 general high school students. As the result of this study, there were no significant differences between the students of the two schools in regards to all the nature of science. However, in the subfields: the observations and the inferences and the changes of scientific theories, the opinions of the general high school students were noticeably higher than those of agricultural specialized high school students. However, the students of the agricultural specialized high school were shown to be more likely to be viewed as experts in the subfields of scientific theories and law. Whereas, in the aspect of the science motivation, the students of the agricultural specialized high schools, showed a meaningfully lower level than those of the general high school. Qualitative analysis of in-depth interviews for 20 focused students of the agricultural specialized high school showed that their low science motivation was caused by their refusal to learn difficult scientific terms, the lack of understanding of the contents of science, and awareness of the relevance between the major and the content of the science curriculum. Thus, to improve their motivation and nature of science itself, the basic science education related to the major is needed. Therefore, it is necessary to improve science text books used in agricultural specialized high school. 본 연구는 농업계 특성화고 학생의 과학 학습 지원 방안을 위한 기초 자료 제시를 목적으로 농업계 특성화고 학생들의 과학의 본성과 과학 동기 수준을 조사하고, 과학과 과학 수업에서 갖는 구체적인 생각을 알아보기 위해 학생들을 심층 면담하였다. 이를 위해 중부지방에 위치한 C농업계 특성화고 2학년 171명, D일반계고 2학년 235명을 대상으로 과학의 본성 검사(SUSSI)와 과학 동기 검사(SMQⅡ)를 실시하였다. 연구결과, 과학의 본성 전체는 농업계 특성화고 학생들과 일반계고 학생 사이에 차이를 보이지 않았다. 그러나 하위 영역인 관찰과 추론, 과학적 이론 변화는 일반계고 학생의 전문가 견해가 높았으며, 과학적 이론과 법칙은 농업계 특성화고 학생들이 전문가 견해가 높았다. 과학 동기에서는 농업계 특성화고 학생이 일반계고 학생 보다 낮은 과학 동기 수준을 보였다. 농업계 특성화고에 재학중인 20명의 초점 학생들을 선별하여 심층 면담을 진행하고, 이를 질적 분석한 결과 농업계 특성화고 학생들은 어려운 과학용어, 부족한 개념 이해 수준, 전공과 과학 교과 내용 사이의 관련성을 인지 하지 못하는 점 등으로 인해 낮은 과학 동기를 보였음을 알 수 있었다. 따라서 농업계 특성화고 학생들의 낮은 과학의 본성 영역을 함양시키고 과학 동기를 높이기 위해 전공과 관련한 기초 과학 교육이 필요하며, 이에 따라 농업계 특성화고에서 사용하는 과학 교과서를 개선할 필요성이 있다.

인공지능을 주제로 한 과학탐구실험 교과 내 ‘첨단과학탐구’ 단원 수업 프로그램의 개발 및 적용

허희정(Heo, Heejung),천재순(Chun, Jaesun) 학습자중심교과교육학회 2022 학습자중심교과교육연구 Vol.22 No.8

목적 본 연구는 고등학교 과학탐구실험 교과의 ‘첨단과학탐구’ 단원에 적용 가능한 인공지능을 주제로 한 수업프로그램을 개발하고 적용하여 과학과 핵심역량 설문과 면담을 통해 프로그램의 효과성을 확인하는 것을 목적으로 한다. 방법 최근 3년간의 뉴스분석을 통해 첨단과학탐구 단원에 적용 가능한 주제로 인공지능을 선정하였고, 총 5차시의 교수-학습 지도안을 개발하였다. 개발된 프로그램의 효과성을 확인하기 위해 단일군 사전, 사후 과학과 핵심역량 평가설문을 실시하였고, SPSS 28.0을 이용하여 기술통계, 대응표본 t-검정, 독립표본 t-검정, 공분산분석을 실시하였다. 또, 수업만족도는 5점 리커드 척도로 설문조사를 실시하였으며 면담을 통해 정량적인 분석을 보완하였다. 결과 개발된 프로그램을 고등학교 1학년 학생들 160명에게 적용한 후 사전, 사후 설문지에 모두 참여한 학생 103명을 대상으로 수업의 효과성 분석을 하였다. 과학과 핵심역량을 대응표본 t-검정 한 결과 과학과 핵심역량 전체와 하위 영역 모두에서 사전, 사후 검사 간에 통계적으로 유의한 차이가 있었다. 또, 공통교과로서의 효과성 분석을 위해 사전에 코딩 관련 프로그래밍을 접해 본 학생들과 접해 보지 못했던 학생들 간 독립표본 t-검정을 시행한 결과 과학과 핵심역량은 유의미한 차이를 보였으나 사전검사를 공변량으로 처리하고 공분산분석을 실시한 사후검사의 결과에서 프로그램이 두 집단의 과학과 핵심역량에 미치는 효과는 통계적으로 차이가 없었다. 즉, 개발된 프로그램은 코딩 관련 사전 지식이 있는 학생들에게만 유의미한 효과를 나타낸 것이 아니라 사전 지식이 없는 학생들에게도 유의미한 효과를 나타내었다. 따라서 본 프로그램은 선택 교육과정 진입 전 형평성을 제공해주는 공통교과의 기능을 해 주었다는 데에 의의가 있다. 또, 개발된 프로그램의 수업만족도는 4.39점으로 높게 나타났다. 결론 개발된 인공지능을 주제로 한 수업 프로그램은 과학과 핵심역량 및 수업 만족도 향상에 효과적이었다. 실생활에서 문제 상황을 찾고 인공지능을 활용하여 해결하기 위한 방안을 모색하는 활동을 통해 학생들의 과학과 핵심역량이 유의미하게 상승되었다. 따라서 본 수업은 고등학교 과학탐구실험 교과 중 ‘첨단과학탐구’ 단원의 수업 프로그램으로 활용될 수 있으며 해당 단원에서의 탐구활동이 다른 단원에 비해 부족했다고 평가되었던 선행연구 결과를 보완할 수 있을 것이다. 앞으로도 과학과 핵심역량을 향상시킬 수 있으면서 동시에 미래 지향적인 교육을 가능하게 하는 첨단 과학 탐구 단원에서의 과학탐구실험 프로그램 개발이 꾸준히 이어지길 기대한다. Objectives The purpose of this study is to develop and apply a class program on the subject of artificial intelligence(AI) that can be applied to the ‘Advanced Science Research’ section of the high school science laboratory experiment course, and to confirm the effectiveness of the program through the questionnaire of the science core competencies and interview. Methods Through news analysis of the last three years, AI was selected as a topic applicable to the ‘Advanced Science Research’ section, and a total of five-period teaching-learning plan was developed. To confirm the effectiveness of the developed program, the evaluation questionnaire of the science core competencies was conducted before and after the application of the program, and the paired sample t-test was conducted, and descriptive statistics, paired-sample t-test, independent sample t-test, and covariance analysis were performed using SPSS 28.0. In addition, a survey was conducted with a 5-point Likeard scale for class satisfaction, and quantitative analysis was supplemented through interviews. Results After applying the developed program to 160 first-year high school students, the effectiveness of classes was analyzed for 103 students who participated in both the pre-test and the post-test in the questionnaires. As a result of the paired sample t-test for science core competencies, there was a statistically significant difference between the pre-test and the post-test in both total science core competencies and their sub-domains. In addition, in order to analyze the effectiveness as a common subject, an independent sample t-test was conducted between students who had previously encountered coding-related programming and those who had not encountered it. There was no statistical difference in the effect of the program on the science core competencies of the two groups in the results of the post-test using covariance analysis. That is, the developed program showed a significant effect not only on students with prior knowledge of coding, but also on students without prior knowledge. Therefore, this program is meaningful in that it served as a common subject that provides equity before entering the optional curriculum. In addition, the class satisfaction of the developed program was high at 4.39 points. Conclusions The developed AI-theme class program was effective in improving science core competencies and class satisfaction. Through the activities of finding problem situations in real life and finding ways to solve them using AI, the students science core competencies were significantly increased. Therefore, this class can be used as a class program for the ‘Advanced Science Research’ section among high school science laboratory experiment course, and it will be able to supplement the results of previous research, which was evaluated as lacking in inquiry activities in that unit compared to other units. It is hoped that the development of science research and experiment programs in the ‘Advanced Science Research’ section will be continued to improve science core competencies while at the same time enabling future-oriented education.

영재교육종합데이터베이스(GED)의 역량중심 영재교육 프로그램 중 중등 생명과학 영역 분석

채지흔(Jiheun Chae),천재순(Jaesun Chun) 학습자중심교과교육학회 2024 학습자중심교과교육연구 Vol.24 No.7

목적 본 연구는 영재교육종합데이터베이스(Gifted Education Database, GED)의 역량중심 영재교육 프로그램 중 중등 생명과학 영역을 분석하여 영재교육 프로그램의 개선을 위한 시사점을 제공하는 것을 목적으로 한다. 방법 GED의 역량중심 영재교육 프로그램 중 중등 생명과학 영역은 2018년의 ‘체제와 상호작용’, 2019년의 ‘에너지’와 ‘안정성’, 2020년의 ‘변화의 패턴’을 대주제로 하여 구성되어 있다. 각각의 대주제는 여러 소주제로 이루어져 있으며 소주제별로 독립적인 수업이 이루어지기 때문에, 하나의 소주제로 묶여 있는 수업을 하나의 개별적인 프로그램으로 간주하여 분석하였다. 프로그램의 분석을 위하여 ‘과학 영재교육 프로그램 평가 기준’을 분석 도구로 사용하였다. 프로그램의 분석에는 영재교육의 경험이 있는 현직 과학 교사 3인이 참여하였으며, 연구의 객관성을 확보하기 위하여 분석 결과를 지속적으로 논의하여 의견을 수렴하는 과정을 거쳤다. 결과 전체 15개의 프로그램의 분석 결과, 평가 기준의 36점 만점에 22점∼30점의 점수가 나타난다. 전체 15개의 프로그램을 평가기준의 주요 특성별로 살펴보면, 15개의 프로그램에서 가장 잘 나타나는 특성은 ‘3. 목표의 명확성’이며, 다음으로는 ‘2. 재능 계발기회’와 ‘5. 평가 요소’, ‘6. 학습 활동’이 그 뒤를 따른다. 반면에, 15개의 프로그램에서 가장 잘 나타나지 않는 특성은 ‘10. 프로그램 구성 요소의 일관성 및 다른 교육자의 사용 편의성’이며, 다음으로는 ‘7. 교수 전략’과 ‘1. 차별화의 본질’이 그 뒤를 따른다. 결론 향후 중등 생명과학 영역의 역량중심 영재교육 프로그램을 개발할 때 ‘10. 프로그램 구성 요소의 일관성 및 다른 교육자의 사용 편의성’과 ‘7. 교수 전략’, ‘1. 차별화의 본질’의 특성을 보완하면 영재 학생이 가진 재능과 특성을 적극적으로 지원할 수 있을 것이며 영재교육 현장에서 영재교육을 담당하는 교사들이 프로그램을 적절히 활용할 수 있을 것이다. 이와 더불어, 초등 생명과학 영역의 역량중심 영재교육 프로그램을 추가적으로 분석하여 중등 영재교육과의 긴밀한 연계가 필요하며, 본 연구에서 분석한 프로그램이 영재교육 현장에서 어떻게 구현되는지 직접 수업을 관찰하는 연구가 추가적으로 이루어질 필요가 있다. Objectives The purpose of this study is to provide implications for improving gifted education programs by analyzing the secondary life science among the competency-based gifted education programs of the Gifted Education Database (GED). Methods Among GED's competency-based gifted education programs, the secondary life science consists of ‘system and interaction’ in 2018, ‘energy’ and ‘stability’ in 2019, and ‘pattern of change’ in 2020. Since each major theme consists of several sub-topics and independent classes are conducted for each sub-topic, the class grouped into one sub-topic was considered as an individual program and analyzed. For the analysis of the program, the ‘evaluation criteria for science gifted education programs’ were used as an analysis tool. Three incumbent science teachers with experience in gifted education participated in the analysis of the program, and in order to secure the objectivity of the research, the analysis results were continuously discussed and opinions were collected. Results As a result of the analysis of a total of 15 programs, a score of 22 to 30 out of 36 points of the evaluation criteria appears. Looking at the main characteristics of the evaluation criteria, the most common characteristics in the 15 programs are ‘3. target clarity’, followed by ‘2. talent development opportunity’, ‘5. evaluation element’, and ‘6. learning activity’. On the other hand, the least visible characteristics in 15 programs are ‘10. consistency of program components and ease of use by other educators’, followed by ‘7. teaching strategy’ and ‘1. nature of differentiation’. Conclusions When developing competency-based gifted education programs in the secondary life science in the future, it is necessary to supplement the characteristics of ‘10. consistency of program components and ease of use by other educators’, ‘7. teaching strategy’, and ‘1. nature of differentiation’. Through this, it will be possible to actively support the talents and characteristics of gifted students, and teachers in the field of gifted education will be able to properly utilize the programs. In addition, it is necessary to further analyze competency-based gifted education programs in the elementary life science to closely link with secondary gifted education. Also, it is necessary to conduct additional research to directly observe classes on how the programs analyzed in this study are implemented in the field of gifted education.

Analysis of Teachers’ Role for CoⅡaboratⅣe CreatⅣity Development

Ye-Ji Jung,Jaesun Chun 한국교원대학교 뇌기반교육연구소 2017 Brain, Digital, & Learning Vol.7 No.2

This study analyzed the factors affecting the creatⅣity development which is covered in the papers on the subject of creatⅣity in Korea and abroad. First, 11 factors and 15 factors for creatⅣity development were extracted from the papers about teachers’ behavior guidelines for creatⅣity development and coⅡaboratⅣe creatⅣity, which is more important in modern society, respectⅣely. It is especiaⅡy important to gⅣe open tasks, to create a free and permissⅣe atmosphere, and to encourage thinking processes through teacher-directed questions. Furthermore, in order to develop coⅡaboratⅣe creatⅣity, it is necessary to recognize the importance of various interactions among members and promote appropriate communication between students and students as weⅡ as teachers and students. The purpose of this study is to explore the direction of school education and the role of teachers to develop students coⅡaboratⅣe creatⅣity.

The Nuclear Localization of Glycogen Synthase Kinase 3beta Is Required Its Putative PY-Nuclear Localization Sequences

신성화,강상순,이은정,JAESUN CHUN,현성희,Youg Il Kim 한국분자세포생물학회 2012 Molecules and cells Vol.34 No.4

Glycogen synthase kinase-3(GSK-3), which is a mem-ber of the serine/threonine kinase family, has been shown to be crucial for cellular survival, differentiation, and metabolism. Here, we present evidence that GSK-3 is associated with the karyopherin 2 (Kap 2) (102-kDa), which functions as a substrate for transportation into the nucleus. A potential PY-NLS motif (109IVRLRYFFY117) was observed, which is similar with the consensus PY NLS motif (R/K/H)X2–5PY in the GSK-3 catalytic domain. Using a pull down approach, we observed that GSK-3 physically interacts with Kap 2 both in vivo and in vitro. Secondly, GSK-3 and Kap 2 were shown to be co-localized by confocal microscopy. The localization of GSK-3 to the nuclear region was disrupted by putative Kap 2 binding site mutation. Furthermore, in transient transfection assays, the Kap 2 binding site mutant induced a substantial reduction in the in vivo serine/threonine phosphorylation of GSK-3, where- as the GSK-3 wild type did not. Thus, our observations indicated that Kap 2 imports GSK-3 through its putative PY NLS motif from the cytoplasm to the nucleus and increases its kinase activity.

Human skeletal dysplasia caused by a constitutive activated transient receptor potential vanilloid 4 (TRPV4) cation channel mutation

강상순,신성화,오정균,JAESUN CHUN 생화학분자생물학회 2012 Experimental and molecular medicine Vol.44 No.12

The transient receptor potential vanilloid 4 (TRPV4)cation channel, a member of the TRP vanilloid subfamily,is expressed in a broad range of tissues where it participates in the generation of Ca2+ signals and/or depolarization of the membrane potential. Regulation of TRPV4 abundance at the cell surface is critical for osmo- and mechanotransduction. Defects in TRPV4are the cause of several human diseases, including brachyolmia type 3 (MIM:113500) (also known as brachyrachia or spondylometaphyseal dysplasia Kozlowski type [MIM:118452]), and metatropic dysplasia (MIM:156530) (also called metatropic dwarfism or parastremmatic dwarfism [MIM:168400]). These bone dysplasia mutants are characterized by severe dwarfism,kyphoscoliosis, distortion and bowing of the extremities,and contractures of the large joints. These diseases are characterized by a combination of decreased bone density, bowing of the long bones, platyspondyly,and striking irregularities of endochondral ossification with areas of calcific stippling and streaking in radiolucent epiphyses, metaphyses, and apophyses. In this review, we discuss the potential effect of the mutation on the regulation of TRPV4 functions,which are related to human diseases through deviated function. In particular, we emphasize how the constitutive active TRPV4 mutant affects endochondral ossification with a reduced number of hypertrophic chondrocytes and the presence of cartilage islands within the zone of primary mineralization. In addition,we summarize current knowledge about the role of TRPV4 in the pathogenesis of several diseases.