請用此 Handle URI 來引用此文件：
http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/92352完整後設資料紀錄
| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 鄭貽生 | zh_TW |
| dc.contributor.advisor | Yi-Sheng Cheng | en |
| dc.contributor.author | 陳健笙 | zh_TW |
| dc.contributor.author | Jian-Sheng Chen | en |
| dc.date.accessioned | 2024-03-21T16:45:18Z | - |
| dc.date.available | 2026-02-28 | - |
| dc.date.copyright | 2024-03-21 | - |
| dc.date.issued | 2024 | - |
| dc.date.submitted | 2024-02-03 | - |
| dc.identifier.citation | 洪珩宸 (2017) 阿拉伯芥 HDA15 組蛋白去乙醯基酶功能區結構及其受磷酸化調控機制之研究。國立臺灣大學植物科學研究所碩士論文。
劉庭君 (2019) 阿拉伯芥組蛋白去乙醯基酶 HDA15 多聚體與酵素活性之關鍵胺基酸研究。國立臺灣大學植物科學研究所碩士論文。 Adasme, M. F., Linnemann, K. L., Bolz, S. N., Kaiser, F., Salentin, S., Haupt, V. J., & Schroeder, M. (2021). PLIP 2021: Expanding the scope of the protein–ligand interaction profiler to DNA and RNA. Nucleic Acids Research, 49(W1), W530-W534. Annunziato, A. T., & Hansen, J. C. (2001). Role of histone acetylation in the assembly and modulation of chromatin structures. Gene Expression The Journal of Liver Research, 9, 37-61. Banta, J. A., & Richards, C. L. (2018). Quantitative epigenetics and evolution. Heredity, 121, 210-224. Berman, H. M., Westbrook, J., Feng, Z., Gilliland, G., Bhat, T. N., Weissig, H., Shindyalov, I. N., & Bourne, P. E. (2000). The protein data bank. Nucleic Acids Research, 28, 235-242. Bernier, G. (1988). The control of floral evocation and morphogenesis. Annual Review of Plant Physiology and Plant Molecular Biology, 39, 175-219. Bernier, G., & Périlleux, C. (2005). A physiological overview of the genetics of flowering time control. Plant Biotechnology Journal, 3, 3-16. Besseau, S., Li, J., & Palva, E. T. (2012). WRKY54 and WRKY70 co-operate as negative regulators of leaf senescence in Arabidopsis thaliana. Journal of Experimental Botany, 63, 2667-2679. Bouché, F., Lobet, G., Tocquin, P., & Périlleux, C. (2016). FLOR-ID: an interactive database of flowering-time gene networks in Arabidopsis thaliana. Nucleic Acids Research, 44(D1), D1167-D1171. Bradford, M. M. (1976). A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Analytical Biochemistry, 72, 248-254. Chen, C.-Y., Tu, Y.-T., Hsu, J.-C., Hung, H.-C., Liu, T.-C., Lee, Y.-H., Chou, C.-C., Cheng, Y.-S., & Wu, K. (2020). Structure of Arabidopsis HISTONE DEACETYLASE15. Plant Physiology, 184, 1585-1600. Chen, H., Lai, Z., Shi, J., Xiao, Y., Chen, Z., & Xu, X. (2010). Roles of Arabidopsis WRKY18, WRKY40 and WRKY60 transcription factors in plant responses to abscisic acid and abiotic stress. BMC Plant Biology, 10, 1-15. Chen, Y.-F., Li, L.-Q., Xu, Q., Kong, Y.-H., Wang, H., & Wu, W.-H. (2009). The WRKY6 transcription factor modulates PHOSPHATE1 expression in response to low Pi stress in Arabidopsis. The Plant Cell, 21, 3554-3566. Ciolkowski, I., Wanke, D., Birkenbihl, R. P., & Somssich, I. E. (2008). Studies on DNA-binding selectivity of WRKY transcription factors lend structural clues into WRKY-domain function. Plant Molecular Biology, 68, 81-92. Crevillen, P., & Dean, C. (2011). Regulation of the floral repressor gene FLC: the complexity of transcription in a chromatin context. Current Opinion in Plant Biology, 14, 38-44. Crevillen, P., Sonmez, C., Wu, Z., & Dean, C. (2013). A gene loop containing the floral repressor FLC is disrupted in the early phase of vernalization. The EMBO Journal, 32, 140-148. Csorba, T., Questa, J. I., Sun, Q., & Dean, C. (2014). Antisense COOLAIR mediates the coordinated switching of chromatin states at FLC during vernalization. Proceedings of the National Academy of Sciences of the United States of America, 111, 16160-16165. DeLano, W. L. (2002). Pymol: An open-source molecular graphics tool. CCP4 Newsl. Protein Crystallogr, 40, 82-92. Deng, W., Ying, H., Helliwell, C. A., Taylor, J. M., Peacock, W. J., & Dennis, E. S. (2011). FLOWERING LOCUS C (FLC) regulates development pathways throughout the life cycle of Arabidopsis. Proceedings of the National Academy of Sciences of the United States of America, 108, 6680-6685. Dhatterwal, P., Basu, S., Mehrotra, S., & Mehrotra, R. (2019). Genome wide analysis of W-box element in Arabidopsis thaliana reveals TGAC motif with genes down regulated by heat and salinity. Scientific Reports, 9, 1681. Ding, Y., Shi, Y., & Yang, S. (2020). Molecular regulation of plant responses to environmental temperatures. Molecular Plant, 13, 544-564. Eswar, N., Eramian, D., Webb, B., Shen, M.-Y., & Sali, A. (2008). Protein structure modeling with MODELLER. Structural Proteomics: High-throughput Methods, 145-159. Eulgem, T., Rushton, P. J., Robatzek, S., & Somssich, I. E. (2000). The WRKY superfamily of plant transcription factors. Trends in Plant Science, 5, 199-206. Fitz-James, M. H., & Cavalli, G. (2022). Molecular mechanisms of transgenerational epigenetic inheritance. Nature Reviews Genetics, 23, 325-341. Franke, D., & Svergun, D. I. (2009). DAMMIF, a program for rapid ab-initio shape determination in small-angle scattering. Journal of Applied Crystallography, 42, 342-346. Glatt, S., Alfieri, C., & Müller, C. W. (2011). Recognizing and remodeling the nucleosome. Current Opinion in Structural Biology, 21, 335-341. Grillo, M. A., Li, C., Hammond, M., Wang, L., & Schemske, D. W. (2013). Genetic architecture of flowering time differentiation between locally adapted populations of Arabidopsis thaliana. New Phytologist, 197, 1321-1331. Grzechowiak, M., Ruszkowska, A., Sliwiak, J., Urbanowicz, A., Jaskolski, M., & Ruszkowski, M. (2022). New aspects of DNA recognition by group II WRKY transcription factor revealed by structural and functional study of AtWRKY18 DNA binding domain. International Journal of Biological Macromolecules, 213, 589-601. He, Y. (2009). Control of the transition to flowering by chromatin modifications. Molecular Plant, 2, 554-564. Heintzman, N. D., Stuart, R. K., Hon, G., Fu, Y., Ching, C. W., Hawkins, R. D., Barrera, L. O., Van Calcar, S., Qu, C., & Ching, K. A. (2007). Distinct and predictive chromatin signatures of transcriptional promoters and enhancers in the human genome. Nature Genetics, 39, 311-318. Heo, J. B., & Sung, S. (2011). Vernalization-mediated epigenetic silencing by a long intronic noncoding RNA. Science, 331, 76-79. Hsin, K.-T., Hsieh, M.-C., Lee, Y.-H., Lin, K.-C., & Cheng, Y.-S. (2022). Insight into the phylogeny and binding ability of WRKY transcription factors. International Journal of Molecular Sciences, 23, 2895. Hu, X., Kong, X., Wang, C., Ma, L., Zhao, J., Wei, J., Zhang, X., Loake, G. J., Zhang, T., & Huang, J. (2014). Proteasome-mediated degradation of FRIGIDA modulates flowering time in Arabidopsis during vernalization. The Plant Cell, 26, 4763-4781. Hu, Y., Zhang, L., He, S., Huang, M., Tan, J., Zhao, L., Yan, S., Li, H., Zhou, K., & Liang, Y. (2012). Cold stress selectively unsilences tandem repeats in heterochromatin associated with accumulation of H3K9ac. Plant, Cell & Environment, 35, 2130-2142. Huang, D., Lan, W., Ma, W., Huang, R., Lin, W., Li, M., Chen, C. Y., Wu, K., & Miao, Y. (2022). WHIRLY1 recruits the histone deacetylase HDA15 repressing leaf senescence and flowering in Arabidopsis. Journal of Integrative Plant Biology, 64, 1411-1429. Humphrey, W., Dalke, A., & Schulten, K. (1996). VMD: visual molecular dynamics. Journal of Molecular Graphics, 14, 33-38. Hung, F.-Y., Shih, Y.-H., Lin, P.-Y., Feng, Y.-R., Li, C., & Wu, K. (2022). WRKY63 transcriptional activation of COOLAIR and COLDAIR regulates vernalization-induced flowering. Plant Physiology, 190, 532-547. Jégu, T., Latrasse, D., Delarue, M., Hirt, H., Domenichini, S., Ariel, F., Crespi, M., Bergounioux, C., Raynaud, C., & Benhamed, M. (2014). The BAF60 subunit of the SWI/SNF chromatin-remodeling complex directly controls the formation of a gene loop at FLOWERING LOCUS C in Arabidopsis. The Plant Cell, 26, 538-551. Khan, M. R. G., Ai, X. Y., & Zhang, J. Z. (2014). Genetic regulation of flowering time in annual and perennial plants. Wiley Interdisciplinary Reviews: RNA, 5, 347-359. Kim, D.-H., & Sung, S. (2017). Vernalization-triggered intragenic chromatin loop formation by long noncoding RNAs. Developmental Cell, 40, 302-312. e304. Kim, D.-H., Xi, Y., & Sung, S. (2017). Modular function of long noncoding RNA, COLDAIR, in the vernalization response. PLoS Genetics, 13, e1006939. Kim, K.-C., Lai, Z., Fan, B., & Chen, Z. (2008). Arabidopsis WRKY38 and WRKY62 transcription factors interact with histone deacetylase 19 in basal defense. The Plant Cell, 20, 2357-2371. Kim, W., Latrasse, D., Servet, C., & Zhou, D.-X. (2013). Arabidopsis histone deacetylase HDA9 regulates flowering time through repression of AGL19. Biochemical and Biophysical Research Communications, 432, 394-398. Kumar, V., Thakur, J. K., & Prasad, M. (2021). Histone acetylation dynamics regulating plant development and stress responses. Cellular and Molecular Life Sciences, 78, 4467-4486. Larkin, M. A., Blackshields, G., Brown, N. P., Chenna, R., McGettigan, P. A., McWilliam, H., Valentin, F., Wallace, I. M., Wilm, A., & Lopez, R. (2007). Clustal W and Clustal X version 2.0. Bioinformatics, 23, 2947-2948. Lee, H. G., & Seo, P. J. (2019). MYB96 recruits the HDA15 protein to suppress negative regulators of ABA signaling in Arabidopsis. Nature Communications, 10, 1713. Lee, J. H., Yoo, S. J., Park, S. H., Hwang, I., Lee, J. S., & Ahn, J. H. (2007). Role of SVP in the control of flowering time by ambient temperature in Arabidopsis. Genes & Development, 21, 397-402. Li, H.-L., Qu, L., Guo, D., Wang, Y., Zhu, J.-H., & Peng, S.-Q. (2020). Histone deacetylase interacts with a WRKY transcription factor to regulate the expression of the small rubber particle protein gene from Hevea brasiliensis. Industrial Crops and Products, 145, 111989. Li, W., Wang, H., & Yu, D. (2016). Arabidopsis WRKY transcription factors WRKY12 and WRKY13 oppositely regulate flowering under short-day conditions. Molecular Plant, 9, 1492-1503. Li, Z., Lathe, R. S., Li, J., He, H., & Bhalerao, R. P. (2022). Towards understanding the biological foundations of perenniality. Trends in Plant Science, 27, 56-68. Liu, C., Xi, W., Shen, L., Tan, C., & Yu, H. (2009). Regulation of floral patterning by flowering time genes. Developmental Cell, 16, 711-722. Madrid, E., Chandler, J. W., & Coupland, G. (2021). Gene regulatory networks controlled by FLOWERING LOCUS C that confer variation in seasonal flowering and life history. Journal of Experimental Botany, 72, 4-14. Mahmood, T., He, S., Abdullah, M., Sajjad, M., Jia, Y., Ahmar, S., Fu, G., Chen, B., & Du, X. (2023). Epigenetic insight into floral transition and seed development in plants. Plant Science, 111926. Manalastas-Cantos, K., Konarev, P. V., Hajizadeh, N. R., Kikhney, A. G., Petoukhov, M. V., Molodenskiy, D. S., Panjkovich, A., Mertens, H. D., Gruzinov, A., & Borges, C. (2021). ATSAS 3.0: expanded functionality and new tools for small-angle scattering data analysis. Journal of Applied Crystallography, 54, 343-355. Martí-Renom, M. A., Stuart, A. C., Fiser, A., Sánchez, R., Melo, F., & Šali, A. (2000). Comparative protein structure modeling of genes and genomes. Annual Review of Biophysics and Biomolecular Structure, 29, 291-325. Mateos, J. L., Madrigal, P., Tsuda, K., Rawat, V., Richter, R., Romera-Branchat, M., Fornara, F., Schneeberger, K., Krajewski, P., & Coupland, G. (2015). Combinatorial activities of SHORT VEGETATIVE PHASE and FLOWERING LOCUS C define distinct modes of flowering regulation in Arabidopsis. Genome Biology, 16, 1-23. Noma, K.-i., Allis, C. D., & Grewal, S. I. (2001). Transitions in distinct histone H3 methylation patterns at the heterochromatin domain boundaries. Science, 293, 1150-1155. Pearson, W. R., & Lipman, D. J. (1988). Improved tools for biological sequence comparison. Proceedings of the National Academy of Sciences of the United States of America, 85, 2444-2448. Qüesta, J. I., Song, J., Geraldo, N., An, H., & Dean, C. (2016). Arabidopsis transcriptional repressor VAL1 triggers Polycomb silencing at FLC during vernalization. Science, 353, 485-488. Quiroz, S., Yustis, J. C., Chávez-Hernández, E. C., Martínez, T., Sanchez, M. d. l. P., Garay-Arroyo, A., Álvarez-Buylla, E. R., & García-Ponce, B. (2021). Beyond the genetic pathways, flowering regulation complexity in Arabidopsis thaliana. International Journal of Molecular Sciences, 22, 5716. Remmert, M., Biegert, A., Hauser, A., & Söding, J. (2012). HHblits: lightning-fast iterative protein sequence searching by HMM-HMM alignment. Nature Methods, 9, 173-175. Ren, X., Chen, Z., Liu, Y., Zhang, H., Zhang, M., Liu, Q., Hong, X., Zhu, J. K., & Gong, Z. (2010). ABO3, a WRKY transcription factor, mediates plant responses to abscisic acid and drought tolerance in Arabidopsis. The Plant Journal, 63, 417-429. Rushton, P. J., Somssich, I. E., Ringler, P., & Shen, Q. J. (2010). WRKY transcription factors. Trends in Plant Science, 15, 247-258. Schneider, R., Bannister, A. J., Myers, F. A., Thorne, A. W., Crane-Robinson, C., & Kouzarides, T. (2004). Histone H3 lysine 4 methylation patterns in higher eukaryotic genes. Nature Cell Biology, 6, 73-77. Schotta, G., Lachner, M., Sarma, K., Ebert, A., Sengupta, R., Reuter, G., Reinberg, D., & Jenuwein, T. (2004). A silencing pathway to induce H3-K9 and H4-K20 trimethylation at constitutive heterochromatin. Genes & Development, 18, 1251-1262. Shen, Y., Lei, T., Cui, X., Liu, X., Zhou, S., Zheng, Y., Guérard, F., Issakidis‐Bourguet, E., & Zhou, D. X. (2019). Arabidopsis histone deacetylase HDA 15 directly represses plant response to elevated ambient temperature. The Plant Journal, 100, 991-1006. Shen, Y., Wei, W., & Zhou, D.-X. (2015). Histone acetylation enzymes coordinate metabolism and gene expression. Trends in Plant Science, 20, 614-621. Shvedunova, M., & Akhtar, A. (2022). Modulation of cellular processes by histone and non-histone protein acetylation. Nature reviews Molecular Cell Biology, 23, 329-349. Sievers, F., Wilm, A., Dineen, D., Gibson, T. J., Karplus, K., Li, W., Lopez, R., McWilliam, H., Remmert, M., & Söding, J. (2011). Fast, scalable generation of high‐quality protein multiple sequence alignments using Clustal Omega. Molecular Systems Biology, 7, 539. Simpson, G. G., & Dean, C. (2002). Arabidopsis, the Rosetta stone of flowering time? Science, 296, 285-289. Song, J., Angel, A., Howard, M., & Dean, C. (2012). Vernalization–a cold-induced epigenetic switch. Journal of Cell Science, 125, 3723-3731. Srikanth, A., & Schmid, M. (2011). Regulation of flowering time: all roads lead to Rome. Cellular and Molecular Life Sciences, 68, 2013-2037. Strange, A., Li, P., Lister, C., Anderson, J., Warthmann, N., Shindo, C., Irwin, J., Nordborg, M., & Dean, C. (2011). Major-effect alleles at relatively few loci underlie distinct vernalization and flowering variation in Arabidopsis accessions. PLoS One, 6, e19949. Swiezewski, S., Liu, F., Magusin, A., & Dean, C. (2009). Cold-induced silencing by long antisense transcripts of an Arabidopsis Polycomb target. Nature, 462, 799-802. Tan-Wong, S. M., Zaugg, J. B., Camblong, J., Xu, Z., Zhang, D. W., Mischo, H. E., Ansari, A. Z., Luscombe, N. M., Steinmetz, L. M., & Proudfoot, N. J. (2012). Gene loops enhance transcriptional directionality. Science, 338, 671-675. Thammasophon, T., Pusadee, T., Bundithya, W., & Naphrom, D. (2023). Effects of Vernalization on Off–Season Flowering and Gene Expression in Sub-Tropical Strawberry cv. Pharachatan 80. Horticulturae, 9, 87. Trabuco, L. G., Villa, E., Mitra, K., Frank, J., & Schulten, K. (2008). Flexible fitting of atomic structures into electron microscopy maps using molecular dynamics. Structure, 16, 673-683. Tu, Y.-T., Chen, C.-Y., Huang, Y.-S., Chang, C.-H., Yen, M.-R., Hsieh, J.-W. A., Chen, P.-Y., & Wu, K. (2022). HISTONE DEACETYLASE 15 and MOS4-associated complex subunits 3A/3B coregulate intron retention of ABA-responsive genes. Plant Physiology, 190, 882-897. Van Aken, O., Zhang, B., Law, S., Narsai, R., & Whelan, J. (2013). AtWRKY40 and AtWRKY63 modulate the expression of stress-responsive nuclear genes encoding mitochondrial and chloroplast proteins. Plant Physiology, 162, 254-271. Vergara, R., Noriega, X., & Pérez, F. J. (2021). VvDAM-SVPs genes are regulated by FLOWERING LOCUS T (VvFT) and not by ABA/low temperature-induced VvCBFs transcription factors in grapevine buds. Planta, 253, 1-10. Volkov, V. V., & Svergun, D. I. (2003). Uniqueness of ab initio shape determination in small-angle scattering. Journal of Applied Crystallography, 36, 860-864. Wriggers, W., & Chacón, P. (2001). Using Situs for the registration of protein structures with low-resolution bead models from X-ray solution scattering. Journal of Applied Crystallography, 34, 773-776. Xu, X., Chen, C., Fan, B., & Chen, Z. (2006). Physical and functional interactions between pathogen-induced Arabidopsis WRKY18, WRKY40, and WRKY60 transcription factors. The Plant Cell, 18, 1310-1326. Yang, H., Howard, M., & Dean, C. (2014). Antagonistic roles for H3K36me3 and H3K27me3 in the cold-induced epigenetic switch at Arabidopsis FLC. Current Biology, 24, 1793-1797. Yoon, H., Seong, G., Lee, S.-J., Lim, C., & Paek, N.-C. (2022). Histone Deacetylases in Rice Development and Stress Responses. Journal of Plant Biology, 65, 175-185. Yu, Y., Liu, Z., Wang, L., Kim, S. G., Seo, P. J., Qiao, M., Wang, N., Li, S., Cao, X., & Park, C. M. (2016). WRKY 71 accelerates flowering via the direct activation of FLOWERING LOCUS T and LEAFY in Arabidopsis thaliana. The Plant Journal, 85, 96-106. Yuan, W., Luo, X., Li, Z., Yang, W., Wang, Y., Liu, R., Du, J., & He, Y. (2016). A cis cold memory element and a trans epigenome reader mediate Polycomb silencing of FLC by vernalization in Arabidopsis. Nature Genetics, 48, 1527-1534. Zeng, X., Gao, Z., Jiang, C., Yang, Y., Liu, R., & He, Y. (2020). HISTONE DEACETYLASE 9 functions with polycomb silencing to repress FLOWERING LOCUS C expression. Plant Physiology, 182, 555-565. Zhang, L., Chen, L., & Yu, D. (2018). Transcription factor WRKY75 interacts with DELLA proteins to affect flowering. Plant Physiology, 176, 790-803. Zhao, L., Peng, T., Chen, C.-Y., Ji, R., Gu, D., Li, T., Zhang, D., Tu, Y.-T., Wu, K., & Liu, X. (2019). HY5 interacts with the histone deacetylase HDA15 to repress hypocotyl cell elongation in photomorphogenesis. Plant Physiology, 180, 1450-1466. Zheng, Y., Ge, J., Bao, C., Chang, W., Liu, J., Shao, J., Liu, X., Su, L., Pan, L., & Zhou, D.-X. (2020). Histone deacetylase HDA9 and WRKY53 transcription factor are mutual antagonists in regulation of plant stress response. Molecular Plant, 13, 598-611. | - |
| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/92352 | - |
| dc.description.abstract | 植物開花時機受到內部和外部因素的精密調控，FLOWERING LOCUS C (FLC) 在抑制植物開花扮演重要的作用，它可抑制FLOWERING LOCUS T (FT) 和SUPPRESSOR OF OVEREXPRESSION OF CONSTANS 1 (SOC1) 等與開花相關的基因表現，抑制開花的發生。WRKY63可促進FLC基因的表現，在過量表現HDA15之阿拉伯芥植株中，FLC的表現量呈現上升趨勢，而質譜分析的初步結果顯示，HDA15與WRKY63可能位於同一複合物中。然而，HDA15和WRKY63之分子結合與功能以及它們如何精細調控FLC基因的表達仍不清楚。在本研究中，蛋白質交互作用實驗pull down assay及Quartz crystal microbalance (QCM) 確認WRKY63藉由DNA結合區 (DB) 與HDA15之去乙醯基催化結構區 (HD) 結合。接著以酵素動力學分析WRKY63對於HDA15之去乙醯基活性的影響，結果顯示，WRKY63與帶有鋅指之HDA15 (HDA15-ZFHD) 結合後會促使HDA15-ZFHD之活性顯著上升。經以fEMSA及QCM對於WRKY63與FLC基因啟動子區域之W box的互動進行分析，結果顯示WRKY63-DB與FLC W1以及FLC W2之親合力較與FLC W3的親和力強度高。以小角度散射方法SAXS結合蛋白質同源結構模擬WRKY63-DB-FLC W1複合體結構，展示了FLC W box中可能與WRKY63結合的關鍵DNA序列保守位點。進一步以QCM和fEMSA分析WRKY63、HDA15及FLC W box的互動。QCM結果顯示，HDA15無法與FLC W box-WRKY63-DB複合體形成三個分子之複合體，並且無法競爭複合體中WRKY63-DB的能力；fEMSA的結果則再次表明HDA15並不會干擾WRKY63-DB與FLC W box結合之能力。本研究深入探討了WRKY63、HDA15及FLC W box之間的功能交互作用關係。將可提供表觀遺傳調控分子HDA15與WRKY63轉錄因子在FLC基因於開花調控分子機制之關鍵訊息。 | zh_TW |
| dc.description.abstract | The timing of plant flowering is intricately regulated by internal and external factors. FLOWERING LOCUS C (FLC) crucially inhibits flowering by suppressing the expression of genes like FLOWERING LOCUS T (FT) and SUPPRESSOR OF OVEREXPRESSION OF CONSTANS 1 (SOC1). WRKY63 enhances FLC gene expression, with Arabidopsis overexpressing HDA15 showing an increased trend in FLC expression. Preliminary mass spectrometry indicates that HDA15 and WRKY63 may form a complex. However, the molecular binding, functions, and fine regulation of FLC gene expression by HDA15 and WRKY63 remain unclear. This study utilized pull-down assays and Quartz Crystal Microbalance (QCM), confirming that WRKY63 binds to HDA15 through its DNA binding domain (DB) and HDA15's deacetylase catalytic structure region (HD). Enzyme kinetics analysis revealed that WRKY63, upon binding to zinc finger-containing HDA15 (HDA15-ZFHD), significantly enhances HDA15-ZFHD's deacetylase activity. Further analysis using fEMSA and QCM on the interaction between WRKY63 and the FLC gene promoter's W box region showed higher affinity of WRKY63-DB for FLC W1 and FLC W2 compared to FLC W3. Small-Angle X-ray Scattering (SAXS) with protein homology structure simulations illustrated the structure of the WRKY63-DB-FLC W1 complex, revealing the conserved DNA binding sites within the WRKY63-DB-FLC W1 complex. QCM and fEMSA analyses investigated WRKY63, HDA15, and FLC W box interactions. HDA15 fails to form a complex with FLC W box-WRKY63-DB and doesn't compete with FLC W box within it, as seen in QCM. fEMSA further verifies HDA15 doesn't disrupt WRKY63-DB binding to FLC W box. This study delves into WRKY63, HDA15, and FLC W box interactions, offering insights into HDA15 and WRKY63 interplay in FLC gene's W box regulation. | en |
| dc.description.provenance | Submitted by admin ntu (admin@lib.ntu.edu.tw) on 2024-03-21T16:45:18Z No. of bitstreams: 0 | en |
| dc.description.provenance | Made available in DSpace on 2024-03-21T16:45:18Z (GMT). No. of bitstreams: 0 | en |
| dc.description.tableofcontents | 論文口試委員會審定書 I
謝辭 II 摘要 III Abstract IV 縮寫對照表 V 目次 VII 圖次 XI 附圖次 XIII 附表次 XIV 第一章、前言 1 1-1. 表觀遺傳學 (epigenetics) 與植物的開花 1 1-2. FLC (FLOWERING LOCUS C) 基因在開花中扮演的角色 2 1-3. 春化作用 (vernalization) 下FLC基因之調控 3 1-4. 春化作用下FLC基因調控之分子機制 3 1-5. AtWRKY63參與FLC基因在春化/非春化下的轉錄調控 5 1-6. 阿拉伯芥WRKY63 5 1-7. 阿拉伯芥HDA15 6 1-8. HDAC是去乙醯基酶 7 1-9. 研究目標 8 第二章、材料與方法 9 2-1. 實驗材料 9 2-2. 實驗方法 9 2-2-1. 質體建構 9 2-2-2. 勝任細胞 (Competent cell) 之製備 10 2-2-3. 轉型作用 (Transformation) 10 2-2-4. HDA15-ZFHD與HDA15-HD蛋白大量表達 10 2-2-5. HDA15-ZFHD與HDA15-HD蛋白純化 11 2-2-6. WRKY63∆C與WRKY63-DB蛋白大量表達 12 2-2-7. WRKY63∆C蛋白純化 12 2-2-8. WRKY63-DB蛋白純化 13 2-2-9. WRKY63-DB蛋白之MBP-tag切除與純化 14 2-2-10. SDS-聚丙烯醯胺膠體電泳 (Sodium dodecyl sulfate polyacrylamide gel electrophoresis, SDS-PAGE) 15 2-2-11. 蛋白質定量分析 15 2-2-12. 西方墨點法 (Western blotting) 16 2-2-13. 粒徑排阻層析分析 (Size exclusion chromatography, SEC) 17 2-2-14. 下拉試驗 (Pull-down assay) 17 2-2-15. 螢光電泳遷移分析 (Fluorescein electrophoretic mobility shift assay, fEMSA) 17 2-2-16. 石英晶體微天平分析蛋白-蛋白之交互作用 18 2-2-17. 去乙醯基酶之酵素動力學檢測 19 2-2-18. 酵素活性檢測 20 2-2-19. 酵素活性影響試驗 21 2-2-20. 小角度X射線散射實驗 (Small Angle X-ray Scattering, SAXS) 21 2-2-21. 蛋白質結構模擬與分子對接 22 第三章、結果 23 3-1. WRKY63蛋白質之純化 23 3-1-1. 全長WRKY63蛋白質在體外(in vitro)呈現非均質穩定態聚集體 (aggregate) 23 3-1-2. 去除WRKY63之Ｃ端可取得均質穩定態之WRKY63-ΔC蛋白質 23 3-1-3. WRKY63-DB蛋白質為單體均質穩定態 25 3-2. HDA15蛋白質之純化 26 3-2-1. HDA15-ZFHD蛋白質之純化 26 3-2-2. HDA15-HD蛋白質之純化 27 3-3. WRKY63與HDA15之交互作用分析 28 3-3-1. WRKY63的DNA結合域與HDA15之去乙醯化催化結構域進行交互作用 28 3-3-2. 利用QCM分析WRKY63與HDA15間之交互作用親和力 28 3-4. HDA15之去乙醯催化活性會受到與WRKY63結合之影響 29 3-5. WRKY63-DB與FLC promoter區域W box之交互作用分析 31 3-5-1. WRKY63-DB可分別與FLC promoter區域的三個W box結合 31 3-5-2. 利用QCM分析WRKY63-DB與FLC promoter區域不同位置之W box結合親和力 32 3-6. WRKY63-DB與FLC W1之結合方式分析 32 3-6-1. 利用SAXS重建WRKY63-DB-FLC W1複合體蛋白質外殼 32 3-6-2. 同源結構重建WRKY63-DB-FLC W1複合體結構 33 3-7. HDA15、WRKY63與W box之結合形式分析 33 3-7-1. 以QCM驗證HDA15不會影響FLC W box-WRKY63-DB複合體之結合 33 3-7-2. 以fEMSA驗證HDA15不會影響WRKY63-DB結合FLC W box 34 第四章、討論 35 4-1. WRKY63蛋白質N端與C端對於WRKY63之穩定性分析 35 4-2. WRKY63之N端促使其形成多聚體 36 4-3. HDA15之去乙醯催化活性受到WRKY63結合之影響 36 4-3-1. WRKY63促使HDA15-ZFHD之活性 (Vmax) 顯著上升 37 4-3-2. WRKY63之N端促使HDA15與受質之親和力下降 (Km上升) 38 4-3-3. WRKY63促成之HDA15-ZFHD活性上升，抵銷WRKY63之N端促成之整體催化效率 (kcat/Km) 降低 39 4-4. FLC W1 DNA序列之正股T11以及反股A18之位置可能是WRKY63的重要結合位點 40 4-5. HDA15不會影響WRKY63與W box之結合 41 第五章、結論 42 參考文獻 44 圖 52 附圖 84 附表 95 | - |
| dc.language.iso | zh_TW | - |
| dc.subject | HDA15 | zh_TW |
| dc.subject | WRKY63 | zh_TW |
| dc.subject | 酵素動力學 | zh_TW |
| dc.subject | 交互作用 | zh_TW |
| dc.subject | FLC W box | zh_TW |
| dc.subject | FLC W box | en |
| dc.subject | interaction | en |
| dc.subject | enzyme kinetics | en |
| dc.subject | HDA15 | en |
| dc.subject | WRKY63 | en |
| dc.title | WRKY63與HDA15之交互作用及分子功能分析 | zh_TW |
| dc.title | Molecular functions and the interplay between WRKY63 and HDA15 | en |
| dc.type | Thesis | - |
| dc.date.schoolyear | 112-1 | - |
| dc.description.degree | 碩士 | - |
| dc.contributor.oralexamcommittee | 吳克強;鄭秋萍;王雅筠;蔡皇龍 | zh_TW |
| dc.contributor.oralexamcommittee | Keqiang Wu;Chiu-Ping Cheng;Ya-Yun Wang;Huang-Lung Tsai | en |
| dc.subject.keyword | WRKY63,HDA15,FLC W box,交互作用,酵素動力學, | zh_TW |
| dc.subject.keyword | WRKY63,HDA15,FLC W box,interaction,enzyme kinetics, | en |
| dc.relation.page | 107 | - |
| dc.identifier.doi | 10.6342/NTU202400408 | - |
| dc.rights.note | 未授權 | - |
| dc.date.accepted | 2024-02-05 | - |
| dc.contributor.author-college | 生命科學院 | - |
| dc.contributor.author-dept | 植物科學研究所 | - |
| 顯示於系所單位： | 植物科學研究所 | |
文件中的檔案：
| 檔案 | 大小 | 格式 | |
|---|---|---|---|
| ntu-112-1.pdf 未授權公開取用 | 6.47 MB | Adobe PDF |
系統中的文件，除了特別指名其著作權條款之外，均受到著作權保護，並且保留所有的權利。