博碩士論文 104826013 詳細資訊




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姓名 廖君宇(Chun-Yu Liao)  查詢紙本館藏   畢業系所 系統生物與生物資訊研究所
論文名稱 利用酵母菌蛋白質體晶片找出與前信使核糖核酸加 工因子19泛素連接?經泛素化作用之受質
(Ubiquitination subtract screening of ubiquitin ligase pre-mRNA-processing factor 19 using yeast proteome microarrays)
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摘要(中) 泛素為一常見於細胞中的小蛋白質,主要用以標記蛋白質受質,並進行後續如去氧
核醣核酸修復、蛋白質水解等路徑。標記泛素的過程稱謂泛素化,其透過泛素活化?、 泛素結合?與泛素連接?將特定受質接上泛素。其中,泛素連接?負責辨識特定的受質, 扮演泛素化中的關鍵角色。前信使核糖核酸加工因子 19 為其中一種泛素連接?,與去 氧核醣核酸修復與信使核糖核酸剪接有所關聯,並被發現在肺癌中有過度表現的現象。 我對大於五千八百種蛋白質的酵母菌蛋白質體晶片進行前信使核糖核酸加工因子 19 的 胞外泛素化實驗,篩選出其對應的四十五個受質,並透過生物資訊的方式找到這四十五 個受質共有結合序列與前信使核糖核酸加工因子 19 透過泛素化受質而進一步產生關聯 的十類生物機制、七類細胞組件、六類分子功能與十二類路徑。另外,在四十五個受質 中,四十個牽涉到七類生物機制、四類細胞組件以及兩類路徑與前信使核糖核酸加工因 子 19 有關聯。此外,有三類生物機制、六類細胞組件和三類分子功能呈現顯著富集。 還有,我找出兩組受質分別有不同的序列模組,提供與前信使核糖核酸加工因子 19 可 能的結合點。根據結果,我找到這些基因本體論與路徑可能透過受質泛素化而被前信使 核糖核酸加工因子 19 所調控,對前信使核糖核酸加工因子 19 進一步的研究方向。
摘要(英) Ubiquitin is a common small protein in cells, it usually served as tag conjugated to the substrate, for DNA repair, proteolysis, and others. This process is called ubiquitination, which ubiquitinates specific substrates by ubiquitin activation enzyme, ubiquitin conjugating enzyme and ubiquitin ligase. Ubiquitin ligase serves as a critical point for substrates specificity. Pre- mRNA-processing factor 19 (PRP19) is one of the ubiquitin ligases, involves in DNA repair, mRNA splicing, and has been found overexpressed in lung cancer. I found forty-five substrates of PRP19 from in vitro ubiquitination assay on yeast proteome chip which contains more than 5800 kinds of the yeast protein. I used bioinformatics analysis to find PRP19 further involving ten classes of biological processes, seven classes of cellular components, six classes of molecule functions, and twelve classes of pathways by ubiquitinates its substrates. Forty of forty-five substrates involved in seven classes of biological processes, four classes of cellular components and two pathways which relate to PRP19.In addition, there were three classes of biological processes, six classes of cellular components, three classes of molecule functions were significantly enriched. Also, I found two groups of substrates had different consensus motifs as putative binding sites with PRP19. According to the result, I found those GO and pathways might be regulated by PRP19-substrates ubiquitination, it gave the direction for
further study on PRP19.
關鍵字(中) ★ 前信使核糖核酸加 工因子19
★ 泛素
★ 泛素化
★ 酵母菌蛋白質體晶片
★ 受質
關鍵字(英) ★ pre-mRNA-processing factor 19
★ ubiquitin
★ Ubiquitination
★ yeast proteome microarrays
★ subtract
論文目次 中文摘要.i
Abstract.ii
Acknowledgments iii
Table of contentsiv
List of figuresvi
List of tables. viii
I. Introduction 1
II. Method10
II.1 Yeast proteome chips production 10
II.2 Ubiquitination enzyme expression and purification12
II.3 Ubiquitination yeast proteome chip assay 12
II.4 Chip image align and data analysis 13
II.5 Bioinformatics analyses 14
II.5.1 Protein annotation information collection14
II.5.2 Gene ontology classification 15
II.5.3 Pathway classification 15
II.5.4 GLAM2 Motif search 16
III. Result 23
III.1 Purification of ubiquitin enzymes23
III.2 Ubiquitination yeast proteome chip assay 24
III.3 Gene ontology classification 24
III.4 Pathway analysis 26
III.5 Motif study 27
IV. Discussion 51
V. Reference 54
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指導教授 陳健生(Chien-Sheng Chen) 審核日期 2017-8-25
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