請用此 Handle URI 來引用此文件：
http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/64325完整後設資料紀錄
| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 沈哲鯤(Che-Kun James Shen) | |
| dc.contributor.author | Chi-Hua Lee | en |
| dc.contributor.author | 李綺華 | zh_TW |
| dc.date.accessioned | 2021-06-16T17:40:49Z | - |
| dc.date.available | 2017-09-18 | |
| dc.date.copyright | 2012-09-18 | |
| dc.date.issued | 2012 | |
| dc.date.submitted | 2012-08-15 | |
| dc.identifier.citation | References
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| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/64325 | - |
| dc.description.abstract | Abstract
The eukaryotic genome is organized into two distinct forms of chromatin, euchromatin and heterochromatin. The entire Drosophila chromosome 4, unlike other autosomes, is almost heterochromatin characterized by methylation of histone 3 at lysine 9 (H3K9me) and the association of heterochromatin protein 1 (HP1) as well as high density of transposable elements (TEs) embedded (Riddle and Elgin 2006). But the 4th chromosome possesses the same density of genes as euchromatin and these genes are actively transcribed during different developmental stages even in the heterochromatic environment (Riddle et al. 2011). However, the regulation mechanism of chromosome 4 genes and TEs is still unclear. This thesis includes two parts. In the first part, we generate null allele of the histone methyltransferase (HMT) dSETDB1, and show that loss of dSETDB1, results in the reduction of H3K9me and HP1-binding on the 4th chromosome. In addition, the binding of POF, a known fourth chromosome-specific protein, and the dSETDB1-controlled H3K9 methylation of this chromosome are interdependent. In the second part, we provide the detail analysis of dSETDB1 null mutant phenotype at high resolution by combining immunostaining images and ChIP-chip analysis data, showing residual H3K9 dimethylation (H3K9me2) and residual binding of HP1 and POF on chromosome 4 in dSETDB1 mutants. The residual H3K9me2 in dSETDB1 mutants depends on SU(VAR)3-9, uncovering the role of SU(VAR)3-9 on chromosome 4. In addition, the distribution of SU(VAR)3-9 along chromosome 4 is dSETDB1-dependent. The major composition of the residual binding regions (RBRs) is transposable elements (TEs) and their expression is derepressed in dSETDB1 mutants. Finally, to determine the effect of dSETDB1 on chromosome 4 gene expression via regulation of HP1-binding, the HP1-binding profiles across chromosome 4 genes were analyzed and the results show that HP1-binding to transcribed gene bodies decrease in dSETDB1 mutants, resulting in down-regulation of gene expression. These results suggest that dSETDB1 enhances transcription by facilitating association of HP1 with transcribed gene bodies. Overall, these observations provide insight into the role of dSETDB1 in regulating chromosome 4 genes and TEs expression. | en |
| dc.description.provenance | Made available in DSpace on 2021-06-16T17:40:49Z (GMT). No. of bitstreams: 1 ntu-101-D92448005-1.pdf: 10277295 bytes, checksum: bc29db9dbdbb9c2ab78d8c4cd94eacd0 (MD5) Previous issue date: 2012 | en |
| dc.description.tableofcontents | Contents
CONTENT…………………………………………………………………………….1 ABSTRACT…………………………………………………………………………...4 中文摘要………………………………………………………………………………6 ABBREVIATIONS........................................................................................................7 CHAPTER I Epigenetic Regulation of Drosophila Chromosome 4 by Histone Methylatransferase dSETDB1…………………………………………………………………………...8 INTRODUCTION…………………………………………………………………...8 Heterochromatin formation……………………………………………………8 Drosophila Chromosome 4…………………………………………………….9 Histone methyltransferase……………………………………………………..10 RESULTS…………………………………………………………………………...13 Generation and characterization of dSETDB1 mutants…………………………13 dSETDB1 is a fourth chromosome-specific histone methyltransferas…………13 HP1- and POF- binding on the polytene fourth chromosome are both dSETDB1 dependent……………………………………………………………………….14 Painting of the fourth chromosome by H3K9me2 is also POF-dependent……..15 DISCUSSION………………………………………………………………………..16 dSETDB1 is an authentic H3K9 methyltransferase regulating the Drosophila Development……………………………………………………………………16 dSETDB1 is mainly a chromosome 4-specific H3K9 methyltransferase in the salivary gland cells, but it also regulates H3K9 methylation globally………….16 Interdependence of dSETDB1, HP1, and POF in the epigenetic painting of chromosome 4…………………………………………………………………..17 Interactive network for epigenetic regulation of the polytene chromosome 4….18 METERIALS AND METHODS……………………………………………………..21 Fly stocks and genetic crosses…………………………………………………..21 Generation and identification of fly lines with FLP-FRT deletions…………….21 Immunostaining of the polytene chromosomes…………………………………22 FIGURES…………………………………………………………………………….23 REFERENCES……………………………………………………………………….29 CHAPTER II Regulation of Heterochromatic Gene Expression by Cooperation of the Histone Methylatransferase dSETDB1 and Su(var)3-9……….………………………………37 ABSTRACT………………………………………………………………………….37 INTRODUCTION……………………………………………………………………39 RESULTS………………………………………………………………………….....42 Mutation of dSETDB1 leaves residual H3K9me2, HP1-binding, and POF- binding on chromosome 4…………………........................................................42 The residual H3K9me2 in dSETDB1 mutants is dependent on SU(VAR)3-9….42 dSETDB1 regulates HP1-binding in heterochromatin………………………….44 The major composition of the residual binding regions is transposable elements…………………………………………………………………………45 The transcription levels of transposable elements increase in dSETDB1 mutants………………………………………………………………………….46 dSETDB1 regulates the distribution of HP1 across chromosome 4 genes……..47 DISCUSSION………………………………………………………………………..49 Su(var)3-9 regulates the residual binding regions in dSETDB1 mutants……….49 dSETDB1 is involved in transposon suppression………………………………51 dSETDB1 regulates chromosome 4 gene expression…………………………..52 METERIALS AND METHODS……………………………………………………53 Fly stocks and genetic crosses…………………………………………………..53 Immunostaining of the polytene chromosomes…………………………………53 Chromatin Imunoprecipitation (ChIP)………………………………………….53 ChIP-chip assays………………………………………………………………..55 Genome tiling array data analysis………………………………………………55 Antibodies………………………………………………………………………56 RNA extraction…………………………………………………………………57 Quantitative PCR………………………………………………………………..57 FIGURES………………………………………………………………………….....58 TABLE……………………………………………………………………………….72 REFERENCES……………………………………………………………………….74 APPENDIX………………………………………………………………………......81 | |
| dc.language.iso | zh-TW | |
| dc.subject | 組蛋白甲基轉移酵素 | zh_TW |
| dc.subject | Histone H3K9 Methyltransferase | en |
| dc.title | 組蛋白甲基轉移酵素dSETDB1調控果蠅第四條染色體的機制研究 | zh_TW |
| dc.title | The Regulatory Mechanisms of Drosophila Chromosome 4 by The Histone H3K9 Methyltransferase dSETDB1 | en |
| dc.type | Thesis | |
| dc.date.schoolyear | 100-2 | |
| dc.description.degree | 博士 | |
| dc.contributor.oralexamcommittee | 謝道時,鍾邦柱,孫以瀚,阮麗蓉,陳瑞華 | |
| dc.subject.keyword | 組蛋白甲基轉移酵素, | zh_TW |
| dc.subject.keyword | Histone H3K9 Methyltransferase, | en |
| dc.relation.page | 81 | |
| dc.rights.note | 有償授權 | |
| dc.date.accepted | 2012-08-15 | |
| dc.contributor.author-college | 醫學院 | zh_TW |
| dc.contributor.author-dept | 分子醫學研究所 | zh_TW |
| 顯示於系所單位： | 分子醫學研究所 | |
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