S-palmitoylation is required for meiotic entry in Schizosaccharomyces pombe

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姓名	范氏青薇(Pham Thi Thanh Vy) 查詢紙本館藏	畢業系所	生命科學系
論文名稱	(S-palmitoylation is required for meiotic entry in Schizosaccharomyces pombe)
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 (2027-8-31以後開放)
摘要(中)	S- palmitoylation是一種轉譯後蛋白質修飾作用，涉及palmitate通過thioester鍵與目標蛋白質的cysteine殘基共價連接。這種修飾促進了這些蛋白質與膜的結合，影響了膜之間的囊泡運輸，並調節了一些蛋白質與蛋白質之交互作用。 S- palmitoylation由一系列跨膜蛋白催化，該蛋白家族包含一個由稱為palmitoyl-acyltransferases (PAT) 的 asp-his-his-cys (DHHC) 組成的催化結構域。儘管S-palmitoylation的基本機制已經被發現一段時間，但PATs與其目標蛋白質的具體相關性仍不清楚。為了更深入地了解 PATs 在生物過程中的作用，我們使用 Schizosaccharomyces pombe作為研究對象，以確定一些 PATs 的受質，並研究驅動這些蛋白質功能的機制。在 S. pombe 中，Akr1、Erf2-Erf4、Pfa3、Pfa5 和 Swf1 屬於 PATs 家族。本研究結果發現，這些蛋白質位於細胞的不同細胞區室，表明PATs酶可能參與了裂殖酵母的各種生物過程。值得注意的是，Erf2-Erf4 和 Akr1 的功能喪失會導致一些減數分裂缺陷，例如無法交配細胞和異常孢子形成。詳細地說，Erf2-Erf4 複合物通過 Ras1 的palmitoylation影響進入減數分裂，Ras1 是一種調節交配費洛蒙反應的基本訊息傳導蛋白。我們還在 Akr1 缺失突變體的 S 期減數分裂中觀察到twin horsetail movement。該結果顯示 Akr1 受質的palmitoylation可能在減數分裂的核配中發揮作用。根據這些發現，我們假設 S- palmitoylation調節了 S. pombe 減數分裂的不同階段。
摘要(英)	S-palmitoylation is a post-translational modification that involves the covalent attachment of palmitate to the cysteine residue of the targeted proteins via a thioester linkage. This modification facilitates the association of these proteins to the membrane and influences vesicular trafficking between membranes as well as regulates some protein-protein interactions. S-palmitoylation is catalyzed by a family of transmembrane proteins that contains a catalytic domain consisting of asp-his-his-cys (DHHC) called palmitoyl-acyltransferases (PATs). Although the basic mechanisms of S-palmitoylation have been discovered for some time, the specific correlations of PATs and their substrates are still unclear. To get more insights into the roles of PATs in biological processes, we used Schizosaccharomyces pombe as the research subject to identify substrates of some PATs as well as investigating the mechanisms that drive the functions of these proteins. In S. pombe, Akr1, Erf2-Erf4, Pfa3, Pfa5, and Swf1 belong to the PATs family. The results of this study have shown that these proteins locate at different cellular compartments of the fission yeast cells, indicating that the PATs enzymes may involve in various biological processes of S. pombe. Notably, the loss of functions of Erf2-Erf4 and Akr1 cause several meiosis defects such as the inability to mate cells and aberrant spore formation. In detail, the Erf2-Erf4 complex affects the meiotic entry by palmitoylation of Ras1, an essential signal transduction protein regulating the mating pheromone response. We also observed a twin horsetail movement at S phase meiosis in Akr1 deleted mutant. This result implies that the palmitoylation of an Akr1′s substrate may play a role in the karyogamy of meiosis. According to these findings, we hypothesize that S-palmitoylation regulates different stages of meiosis in S. pombe.
關鍵字(中)	★ palmitoylation	關鍵字(英)
論文目次	中文摘要 i ABSTRACT ii Acknowledgments iii Table of Contents iv List of Figures vi List of Tables vii Abbreviations viii Chapter I. INTRODUCTION 1 1.1. The structure of PATs 1 1.2. The mechanism of S-palmitoylation 2 1.3. The roles of S-palmitoylation in cellular processes. 4 1.4. Palmitoylation and human disease 4 1.4.1. Huntington’s disease 5 1.4.2. Human cancer 5 1.5. PATs and palmitoylated proteins in S. pombe 6 1.6. Meiosis in S. pombe 7 Chapter II. MATERIAL AND METHOD 10 2.1. Fission yeast strains and growth condition 10 2.2. Lithium acetate transformation 11 2.3. PATs genes deletion and tag fusion 11 2.4. Colony PCR from yeast cells 12 2.5. Gene overexpression 12 2.6. Site-directed mutagenesis 14 2.7. Transformation of E. coli 15 2.8. Crosses and tetrad dissection 15 2.9. Microscopy 15 2.10. Protein extraction and Western blotting 15 Chapter III. RESULT 17 3.1. The PATs family of S. pombe 17 3.2. The involvement of the PATs in the fission yeast meiosis 19 3.3. The role of Erf2-Erf4 in matting of fission yeast 21 3.4. Akr1 is important for karyogamy of meiosis in fission yeast. 24 Chapter IV. CONCLUSION AND DISCUSSION 26 Reference 31
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指導教授	王紹文葉淑丹(Shao-Win Wang Shu-Dan Yeh)	審核日期	2021-8-9
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