水稻熱休克蛋白質OsHSP16.9A與OsHSP101之交互作用分析

以作者查詢圖書館館藏

、以作者查詢臺灣博碩士

、以作者查詢全國書目

、勘誤回報

、線上人數：19

、訪客IP：18.188.40.207

姓名

黃信傑(Xin-Jie Huang) 查詢紙本館藏

畢業系所

生命科學系

論文名稱

水稻熱休克蛋白質OsHSP16.9A與OsHSP101之交互作用分析
(Analysis of the interplay between heat shock proteins OsHSP16.9A and OsHSP101 in rice)

相關論文

★ 第三群LEA蛋白質表現與功能分析	★ 水稻小分子量熱休克蛋白質Oshsp16.9A之N端區域功能性分析
★ 植物逆境蛋白質基因啟動子與功能分析	★ 植物受溫度調控之基因的功能與機制分析
★ 錯誤褶疊蛋白質誘導之擬熱休克反應機制之探討	★ 受熱與ABA調控水稻基因-OsRZFP1之生理功能分析
★ 受熱與ABA調控基因AtRZFP33之生理功能分析	★ 水稻第一族小分子量熱休克蛋白質OsHSP16.9A及OsHSP18.0之生理功能分析
★ 植化物紫草素在小鼠皮膚上增加血管通透性之研究	★ 蝴蝶蘭開花相關基因PaCOL2啟動子之特性分析
★ 利用水稻HSP17.3啟動子探討阿拉伯芥熱休克因子在逆境下對細胞內蛋白質反應之角色分析	★ 蝴蝶蘭開花相關基因PaCOL1 啟動子之特性分析
★ 分析水稻 RING 鋅手指蛋白質 OsRZFP34 與其正向調控蛋白質之交互作用	★ 水稻小分子量熱休克蛋白質- OsHSP16.9A在水稻種子耐熱性之功能分析
★ Oryzasin 1 在水稻種子耐熱性之功能分析	★ 水稻小分子量熱休克蛋白質—OsHSP16.9A關鍵胺基酸分析

檔案

[Endnote RIS 格式]

[Bibtex 格式]

[相關文章]

[文章引用]

[完整記錄]

[館藏目錄]

至系統瀏覽論文 ( 永不開放)

摘要(中)

小分子熱休克蛋白質是植物體內種類相當豐富的一群蛋白質，能與變性蛋白質結合，防止其沉澱而減緩對細胞的傷害。實驗室先前發現OsHSP16.9A與OsHSP101可能在高溫下產生交互作用，也發現OsHSP16.9A與OsHSP101會累積於胚中，並在泡水後兩天開始減少。為了更進一步證實OsHSP16.9A與OsHSP101之交互作用，我們利用比例式雙螢光分子互補系統（rBiFC）來偵測彼此間在細胞內的交互作用。結果顯示OsHSP16.9A與OsHSP101可以在正常及高溫條件下具有交互作用。我們也發現當OsHSP16.9A第74個胺基酸從Glu突變成Asp時，其與OsHSP101的交互作用就會大幅下降。由此可知，OsHSP16.A的第74個胺基酸對於與OsHSP101形成交互作用十分重要。此外，我們也分析了TNG67、CO39、N22、IR64及IR50等五種水稻在種子充實期間OsHSP16.9A與OsHSP101的累積情形。結果顯示OsHSP16.9A及OsHSP101的累積趨勢相當一致，從乳熟期開始累積，並在黃熟期、完熟期達到高峰。綜上所述，水稻會在種子發育期間累積大量OsHSP16.9A及OsHSP101，可能藉此使種子獲得高溫耐受性，直到萌芽後開始衰退，因此，我們認為OsHSP16.9A與OsHSP101間可能有著類似於OsHSA32與OsHSP101間的正回饋關係。

摘要(英)

Small heat shock proteins represent the most abundant heat shock proteins in plants. These proteins have chaperon activity, which can prevent cellular proteins from thermal-induced irreversible denaturation under heat stress. By the proteomic approach, we found that OsHSP101 is a potential candidate interacting with OsHSP16.9A during high temperature. In addition, our previous study found the abundance of OsHSP16.9A and OsHSP101 was in the embryo and vanished after imbibition for two days. To clear up the interplay between OsHSP16.9A and OsHSP101, we used ratiometric bimolecular fluorescence complementation (rBiFC) to investigate the interaction between OsHSP16.9A and OsHSP101. The results show that OsHSP16.9A interacts with OsHSP101 under non-stress and heat stress conditions. Besides, we found that mutation of the 74th amino acid residue, Glu to Asp, of the OsHSP16.9A leads to abolish the interaction of OsHSP16.9A and OsHSP101. Our results indicate that the 74th amino acid of OsHSP16.9A is critical for OsHSP16.9A–OsHSP101 interaction under thermal stress. Besides, we also analyzed OsHSP16.9A and OsHSP101 accumulation level during seed maturation in 5 different rice cultivar, including TNG67, CO39, N22, IR50 and IR28. Accumulation levels of OsHSP16.9A and OsHSP101 showed similar patterns during seed maturation. OsHSP16.9A and OsHSP101 start to accumulate at the milk stage and reach the peak at the hard dough stage or mature stage. These results indicate that accumulation of OsHSP16.9A and OsHSP101 during seed development may be important for seeds to gain thermotolerance. When a suitable environment manifests for seed germination, OsHSP16.9A and OsHSP101 will start to decline. Therefore, we suppose there has a positive feedback loop between OsHSP16.9A and OsHSP101, which similar to OsHSA32 and OsHSP101.

關鍵字(中)

★ 水稻
★ 熱休克蛋白質16.9A
★ 熱休克蛋白質101
★ 交互作用

關鍵字(英)

★ rice
★ OsHSP16.9A
★ OsHSP101
★ interaction

論文目次

摘要 i
Abstract ii
誌謝 iii
圖目錄 v
附錄目錄 vi
縮寫對照表 vii
序論 1
研究起源與目的 7
材料與方法 10
結果 30
討論 36
參考文獻 42
圖表 47
附錄 62

參考文獻

1. Osmond C.B., et al., Stress physiology and the distribution of plants. BioScience. 37, 38-48, (1987).
2. Smirnoff N., Plant stress physiology. eLS, (2014).
3. Mehdy M.C., Active oxygen species in plant defense against pathogens. Plant Physiology. 105, 467, (1994).
4. Serrano R., et al., A glimpse of the mechanisms of ion homeostasis during salt stress. Journal of Experimental Botany, 1023-1036, (1999).
5. Zhu J.-K., Plant salt tolerance. Trends in Plant Science. 6, 66-71, (2001).
6. Suzuki N., et al., ABA is required for plant acclimation to a combination of salt and heat stress. PLoS One. 11, e0147625, (2016).
7. Kawakatsu T., et al., Reducing rice seed storage protein accumulation leads to changes in nutrient quality and storage organelle formation. Plant Physiology. 154, 1842-1854, (2010).
8. Fahad S., et al., Crop production under drought and heat stress: Plant responses and management options. Frontiers in Plant Science. 8, 1147, (2017).
9. Wang W., et al. Biotechnology of plant osmotic stress tolerance physiological and molecular considerations. in IV International Symposium on In Vitro Culture and Horticultural Breeding 560. 2000.
10. Hatfield J.L. and Prueger J.H., Temperature extremes: effect on plant growth and development. Weather Climate Extremes. 10, 4-10, (2015).
11. Wahid A., et al., Heat tolerance in plants: an overview. Environmental experimental botany. 61, 199-223, (2007).
12. Vierling E., The roles of heat shock proteins in plants. Annual Review of Plant Biology. 42, 579-620, (1991).
13. Hong S.-W. and Vierling E., Mutants of Arabidopsis thaliana defective in the acquisition of tolerance to high temperature stress. Proceedings of the National Academy of Sciences. 97, 4392-4397, (2000).
14. Larkindale J., et al., Heat stress phenotypes of Arabidopsis mutants implicate multiple signaling pathways in the acquisition of thermotolerance. Plant Physiology. 138, 882-897, (2005).
15. Rukmini M., D′souza B., and D′souza V., Superoxide dismutase and catalase activities and their correlation with malondialdehyde in schizophrenic patients. Indian Journal of Clinical Biochemistry. 19, 114, (2004).
16. Shigeoka S., et al., Regulation and function of ascorbate peroxidase isoenzymes. Journal of Experimental Botany. 53, 1305-1319, (2002).
17. Savicka M. and Škute N., Effects of high temperature on malondialdehyde content, superoxide production and growth changes in wheat seedlings (Triticum aestivum L.). Ekologija. 56, 26-33, (2010).
18. Adachi M., et al., Oxidative stress impairs the heat stress response and delays unfolded protein recovery. PLoS One. 4, e7719, (2009).
19. Kotak S., et al., Complexity of the heat stress response in plants. Curr Opin Plant Biol. 10, 310-316, (2007).
20. Swindell W.R., Huebner M., and Weber A.P., Transcriptional profiling of Arabidopsis heat shock proteins and transcription factors reveals extensive overlap between heat and non-heat stress response pathways. BMC Genomics. 8, 125, (2007).
21. Schöffl F., Prandl R., and Reindl A., Molecular responses to heat stress, Molecular responses to cold, drought, heat and salt stress in higher plants. 5, 83-97, (1999).
22. Waters E.R., The evolution, function, structure, and expression of the plant sHSPs. Journal of Experimental Botany. 64, 391-403, (2013).
23. de Jong W.W., Leunissen J.A., and Voorter C., Evolution of the alpha-crystallin/small heat-shock protein family. Molecular Biology Evolution. 10, 103-126, (1993).
24. de Jong W.W., Caspers G.J., and Leunissen J.A., Genealogy of the α-crystallin—small heat-shock protein superfamily. International Journal of Biological Macromolecules. 22, 151-162, (1998).
25. Horwitz J., Alpha-crystallin can function as a molecular chaperone. Proceedings of the National Academy of Sciences. 89, 10449-10453, (1992).
26. Haslbeck M., et al., Some like it hot: the structure and function of small heat-shock proteins. Nature Structural Molecular Biology Evolution. 12, 842-846, (2005).
27. Jaya N., Garcia V., and Vierling E., Substrate binding site flexibility of the small heat shock protein molecular chaperones. Proceedings of the National Academy of Sciences. 106, 15604-15609, (2009).
28. Fu X., et al., Small heat shock protein IbpB acts as a robust chaperone in living cells by hierarchically activating its multi-type substrate-binding residues. Journal of Biological Chemistry. 288, 11897-11906, (2013).
29. Basha E., Friedrich K.L., and Vierling E., The N-terminal arm of small heat shock proteins is important for both chaperone activity and substrate specificity. Journal of Biological Chemistry. 281, 39943-39952, (2006).
30. Ungelenk S., et al., Small heat shock proteins sequester misfolding proteins in near-native conformation for cellular protection and efficient refolding. Nature Communications. 7, 1-14, (2016).
31. Basha E., et al., An unusual dimeric small heat shock protein provides insight into the mechanism of this class of chaperones. Journal of Molecular Biology. 425, 1683-1696, (2013).
32. Bepperling A., et al., Alternative bacterial two-component small heat shock protein systems. Proceedings of the National Academy of Sciences. 109, 20407-20412, (2012).
33. Leroux M.R., et al., Structure-function studies on small heat shock protein oligomeric assembly and interaction with unfolded polypeptides. Journal of Biological Chemistry. 272, 24646-24656, (1997).
34. Waters E.R., Lee G.J., and Vierling E., Evolution, structure and function of the small heat shock proteins in plants. Journal of Experimental Botany. 47, 325-338, (1996).
35. Nover L., Scharf K., and Neumann D., Formation of cytoplasmic heat shock granules in tomato cell cultures and leaves. Molecular and Cellular Biology. 3, 1648-1655, (1983).
36. Wang W., et al., Role of plant heat-shock proteins and molecular chaperones in the abiotic stress response. Trends in Plant Science. 9, 244-252, (2004).
37. Bukau B. and Horwich A.L., The Hsp70 and Hsp60 chaperone machines. Cell. 92, 351-366, (1998).
38. Hartl F.U. and Hayer-Hartl M., Molecular chaperones in the cytosol: From nascent chain to folded protein. Science. 295, 1852-1858, (2002).
39. Santhanagopalan I., et al., It takes a dimer to tango: Oligomeric small heat shock proteins dissociate to capture substrate. Journal of Biological Chemistry. 293, 19511-19521, (2018).
40. Wehmeyer N. and Vierling E., The expression of small heat shock proteins in seeds responds to discrete developmental signals and suggests a general protective role in desiccation tolerance. Plant Physiology. 122, 1099-1108, (2000).
41. Haslbeck M. and Vierling E., A first line of stress defense: small heat shock proteins and their function in protein homeostasis. Journal of Molecular Biology. 427, 1537-1548, (2015).
42. Löw D., et al., Cytosolic heat-stress proteins Hsp17.7 class I and Hsp17.3 class II of tomato act as molecular chaperones in vivo. Planta. 211, 575-582, (2000).
43. Ju Y., et al., Overexpression of OsHSP18.0-CI enhances resistance to bacterial leaf streak in rice. Rice. 10, 12, (2017).
44. Kirstein J., et al., Adapting the machine: adaptor proteins for Hsp100/Clp and AAA+ proteases. Nature Reviews Microbiology. 7, 589-599, (2009).
45. Weibezahn J., et al., Thermotolerance requires refolding of aggregated proteins by substrate translocation through the central pore of ClpB. Cell. 119, 653-665, (2004).
46. Michalska K., et al., Structure of Calcarisporiella thermophila Hsp104 disaggregase that antagonizes diverse proteotoxic misfolding events. Structure. 27, 449-463. e447, (2019).
47. Parsell D.A., et al., Protein disaggregation mediated by heat-shock protein Hsp104. Nature. 372, 475-478, (1994).
48. Lee S., et al., The structure of ClpB: a molecular chaperone that rescues proteins from an aggregated state. Cell. 115, 229-240, (2003).
49. Panaretou B. and Zhai C., The heat shock proteins: their roles as multi-component machines for protein folding. Fungal Biology Reviews. 22, 110-119, (2008).
50. Żwirowski S., et al., Hsp70 displaces small heat shock proteins from aggregates to initiate protein refolding. The EMBO journal. 36, 783-796, (2017).
51. Queitsch C., et al., Heat shock protein 101 plays a crucial role in thermotolerance in Arabidopsis. The Plant Cell. 12, 479-492, (2000).
52. Hong S.W., Lee U., and Vierling E., Arabidopsis hot mutants define multiple functions required for acclimation to high temperatures. Plant Physiology. 132, 757-767, (2003).
53. Gurley W.B., Hsp101: A key component for the acquisition of thermotolerance in plants. The Plant Cell. 12, 457-460, (2000).
54. McLoughlin F., et al., Class I and II small heat shock proteins together with HSP101 protect protein translation factors during heat stress. Plant Physiology. 172, 1221-1236, (2016).
55. McLoughlin F., et al., HSP101 interacts with the proteasome and promotes the clearance of ubiquitylated protein aggregates. Plant Physiology. 180, 1829-1847, (2019).
56. Wang W., et al., Genomic variation in 3,010 diverse accessions of Asian cultivated rice. Nature. 557, 43-49, (2018).
57. Oka H.I., Origin of cultivated rice. (2012): Elsevier.
58. Khush G.S., Origin, dispersal, cultivation and variation of rice. Plant Molecular Biology. 35, 25-34, (1997).
59. Casartelli A., et al., Exploring traditional aus-type rice for metabolites conferring drought tolerance. Rice. 11, 9, (2018).
60. Schatz M.C., et al., Whole genome de novo assemblies of three divergent strains of rice, Oryza sativa, document novel gene space of aus and indica. Genome Biology. 15, 506, (2014).
61. Singh U., Aromatic rices. (2000): Int. Rice Res. Inst.
62. Xu X., et al., Resequencing 50 accessions of cultivated and wild rice yields markers for identifying agronomically important genes. Nature Biotechnology. 30, 105, (2012).
63. Baker J., Allen Jr L., and Boote K., Temperature effects on rice at elevated CO2 concentration. Journal of Experimental Botany. 43, 959-964, (1992).
64. Peng S., et al., Rice yields decline with higher night temperature from global warming. Proceedings of the National Academy of Sciences. 101, 9971-9975, (2004).
65. Welch J.R., et al., Rice yields in tropical/subtropical Asia exhibit large but opposing sensitivities to minimum and maximum temperatures. Proceedings of the National Academy of Sciences. 107, 14562-14567, (2010).
66. Yao F., et al., Assessing the impacts of climate change on rice yields in the main rice areas of China. Climatic Change. 80, 395-409, (2007).
67. Sarkar N.K., Kim Y.K., and Grover A., Rice sHsp genes: genomic organization and expression profiling under stress and development. BMC Genomics. 10, 393, (2009).
68. Guan J.C., et al., Characterization of the genomic structures and selective expression profiles of nine class I small heat shock protein genes clustered on two chromosomes in rice (Oryza sativa L.). Plant Molecular Biology. 56, 795-809, (2004).
69. 劉依欣，「水稻第一族小分子量熱休克蛋白質OsHSP16.9A及OsHSP18.0之生理功能分析」，國立中央大學，碩士論文，第1-89頁，(2011)
70. 鄭榮儀，「水稻小分子熱休克蛋白質OsHSP16.9A在水稻種子耐熱性之功能分析」，國立中央大學，碩士論文，第1-107頁，(2015)
71. Grefen C. and Blatt M.R., A 2in1 cloning system enables ratiometric bimolecular fluorescence complementation (rBiFC). Biotechniques. 53, 311-314, (2012).
72. Bradford M.M., 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, (1976).
73. 李易諠，「水稻小分子量熱休克蛋白質Oshsp16.9A之N端區域功能性分析」，國立中央大學，碩士論文，第1-45頁，(2006)
74. Yeh C.H., Chen Y.M., and Lin C.Y., Functional regions of rice heat shock protein, Oshsp16. 9, required for conferring thermotolerance in Escherichia coli. Plant Physiology. 128, 661-668, (2002).
75. 曹雅君, et al., 水稻品种休眠特性的研究. 南京农业大学学报. 24, 1-5, (2001).
76. 林冠甫，「Structure-function relationship of a rice small heat shock protein OsHsp16.9A」，國立中央大學，博士論文，第1-47頁，(2019)
77. Rutsdottir G., et al., Structural model of dodecameric heat-shock protein Hsp21: Flexible N-terminal arms interact with client proteins while C-terminal tails maintain the dodecamer and chaperone activity. Journal of Biological Chemistry. 292, 8103-8121, (2017).
78. Giese K.C., et al., Evidence for an essential function of the N-terminus of a small heat shock protein in vivo, independent of in vitro chaperone activity. Proceedings of the National Academy of Sciences. 102, 18896-18901, (2005).
79. McDonald E.T., et al., Sequence, structure, and dynamic determinants of Hsp27 (HspB1) equilibrium dissociation are encoded by the N-terminal domain. Biochemistry. 51, 1257-1268, (2012).
80. Lin M.y., et al., A positive feedback loop between heat shock protein 101 and heat stress-associated 32-kd protein modulates long-term acquired thermotolerance illustrating diverse heat stress responses in rice varieties. Plant Physiology. 164, 2045-2053, (2014)

指導教授

葉靖輝(Ching-Hui Yeh)

審核日期

2020-6-18

推文