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Graduate School of Ajou University
Department of Neuroscience and Technology Course
3. Theses(Master)
사람 tyrosine hydroxylase 유전자의 발현에서의 Nurr1에의한 세포 특이적인 조절작용
| DC Field | Value | Language |
|---|---|---|
| dc.contributor.advisor | 이명애 | - |
| dc.contributor.author | 서지선 | - |
| dc.date.accessioned | 2019-10-21T06:47:32Z | - |
| dc.date.available | 2019-10-21T06:47:32Z | - |
| dc.date.issued | 2006-02 | - |
| dc.identifier.other | 1206 | - |
| dc.identifier.uri | https://dspace.ajou.ac.kr/handle/2018.oak/16750 | - |
| dc.description | 학위논문(석사)--아주대학교 일반대학원 :신경과학기술과정,2006. 2 | - |
| dc.description.tableofcontents | TABLE OF CONTENTS ABSTRACT ………………………………………………………………… 4 TABLE OF CONTENTS …………………………………………………..... 5 LIST OF FIGURES ……………………………………………………….... 7 I. INTRODUCTION …………………………………………………….…. 8 II. MATERIAL AND METHODS ……….…………………………….…… 11 1. Cell culture ……………………...….…………………………….......... 11 2. Plasmid constructs ............................………………………………….. 11 3. Transient Transfection …………….…………….…………..………… 12 4. Luciferase assay ………………….…………………….…...………… 13 5. Biotinylated Oligonucleotide-streptavidin Pull –Down …..….….…..... 13 6. Western blot analysis …………….………….……………………….... 14 7. Two-dimensional PAGE ………………………………………………. 15 8. Protein visualization and image analysis ………………………...…… 16 9. In-gel digestion ……..............………………………………......…….. 17 10. Mass spectrometry and database search …………………..………....... 18 III. RESULTS ……………………….…………………………..……….... 19 1. Differential regulation of human TH gene by Nurr1 in cell-type dependent Manner ………….………………………...………………… 19 2. The upstream promoter region of the hTH gene, mediating full responsiveness to transcriptional regulation by Nurr1, contains multiple NBRE-like sequence motif ……..…………………………….. 21 3. NBRE-A region is critical for transregulation of the TH promoter Activity by Nurr1 …………………………………………….…………. 22 4. HDAC- independent inhibition of Nurr1-mediated hTH promoter transcriptional repression in F3 cell line ……..…………………………. 24 5. Purification of Nurr1 and recruitment cofactor complex in NBRE-A region ………..……………………………..………..…….... 25 6. Protein differentially expressed in human neural stem cells and human neuroblastoma cells by Nurr1 ………………………………...…. 26 IV. DICUSSION ………….……………………...………………………… 28 V. REFERENCES ….……………………..……………………………….. 42 국문요약 …………………………………….……………………………. 47 Acknowledgement …………………………………………………………. 48 | - |
| dc.language.iso | eng | - |
| dc.publisher | The Graduate School, Ajou University | - |
| dc.rights | 아주대학교 논문은 저작권에 의해 보호받습니다. | - |
| dc.title | 사람 tyrosine hydroxylase 유전자의 발현에서의 Nurr1에의한 세포 특이적인 조절작용 | - |
| dc.title.alternative | Ji Seon Seo | - |
| dc.type | Thesis | - |
| dc.contributor.affiliation | 아주대학교 일반대학원 | - |
| dc.contributor.alternativeName | Ji Seon Seo | - |
| dc.contributor.department | 일반대학원 신경과학기술과정 | - |
| dc.date.awarded | 2006. 2 | - |
| dc.description.degree | Master | - |
| dc.identifier.localId | 565034 | - |
| dc.identifier.url | http://dcoll.ajou.ac.kr:9080/dcollection/jsp/common/DcLoOrgPer.jsp?sItemId=000000001206 | - |
| dc.description.alternativeAbstract | Nuclear receptors represent a large family of transcriptional factors involved in development, differentiation, homeostasis, and cancer. Nurr1, a member of the orphan nuclear receptor superfamily, was recently demonstrated to be of critical importance in the developing central nervous system, where it is required for the generation of midbrain dopaminergic cell. Although its role during dopaminergic neurogenesis has been extensively studied, its transcriptional regulation of TH, the rate-limiting enzyme of dopamine biosynthesis, is not well understood. Here we show that Nurr1 directly mediated transcriptional activation or repression of TH gene through NBRE-A site and a key regulating complex of TH expression, that may be formed in NBRE-A site during dopaminergic neurogenesis. In recent years, a growing number of cofactor has been discovered that participate in the regulation of the transcriptional activity of promoter. So, we present the identification of a cofactors binding in and around NBRE-A, which differentially regulates the transcriptional activity of the human TH promoter in human neural stem cell, F3, and human neuroblastoma cell, SH-SY5Y, we are preformed 2-D gel electrophoresis and LC peptide sequencing analysis. Together, these data suggests that Nurr1 and cofactors may regulate differentially transcription activity of TH promoter in neural stem cell and dopaminergic neurons. Keywords: tyrosine hydroxylase, Nurr1, doapminergic neuron, neural stem cell, neuroblastoma cell. | - |
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