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研究生：	黃柏維 Huang, Po-Wei
論文名稱：	秀麗隱桿線蟲的核仁蛋白 NOL-58 會調控核仁監視反應和 p38 MAPK 途徑藉此對抗細菌感染 Nucleolar protein NOL-58 regulates nucleolar surveillance responses and the p38 MAPK pathway against bacterial infection in C. elegans
指導教授：	陳柏齡 Chen, Po-Lin
學位類別：	碩士 Master
系所名稱：	醫學院 - 微生物及免疫學研究所 Department of Microbiology & Immunology
論文出版年：	2023
畢業學年度：	111
語文別：	英文
論文頁數：	68
中文關鍵詞：	核仁蛋白、NOL-58 、秀麗隱桿線蟲、達卡產氣單胞菌、p38 MAPK 訊息傳遞路徑、先天免疫、核仁監視反應
外文關鍵詞：	nucleolar protein, NOL-58, Caenorhabditis elegans, Aeromonas dhakensis, p38 MAPK, innate immunity, nucleolar surveillance response
相關次數：	點閱：74 下載：0
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Abstract I
中文摘要 II
致謝 III
Introduction 1
1.	Aeromonas dhakensis 1
2.	Caenorhabditis elegans 3
3.	Physiologic of C. elegans 5
4.	Innate immunity of C. elegans 5
5.	Nucleolar proteins 6
6.	NOL-58 8
Materials and Methods 10
1.	Caenorhabditis elegans strains and mutant alleles 10
2.	Bacteria strains and plasmids 10
3.	Media and chemical 11
4.	Cultivation of C. elegans and bleaching 11
5.	RNAi screening of the nucleolar proteins 11
6.	Infectious survival assay of nematodes 12
7.	Measurement of phenotypic and physiologic characteristics of C. elegans 12
8.	Tissue-specific knockdown and epistasis analysis 13
9.	p38 phosphorylation western blot analysis 13
10.	Quantification of nol-58 gene and pmk-1 downstream antimicrobial genes expression 14
11.	The nucleolar surveillance response (NSR) analysis and confocal microscope 15
12.	The construction of the nol-58 overexpression animal 15
13.	Statistical analysis 15
Result 16
1.	Screening of the nucleolar proteins list of C. elegans 16
2.	Worms with knockdown of nol-58 are resistant to A. dhakensis 17
3.	Knockdown of nol-58 leads to phenotypic and physiologic changes in C. elegans 17
4.	Silencing nol-58 expression also mitigates several bacterial infections 19
5.	nol-58 has its effect in neurons, intestine, and germline 19
6.	NOL-58 affects the p38 MPAK innate immunity pathway of C. elegans 20
7.	Expression level of the phosphorylated p38 and downstream antimicrobial genes were upregulated when nol-58 was interfered 22
8.	Interference of nol-58 circumvents the nucleolar stress responses in C. elegans 23
Conclusion 25
Discussion 26
1.	Functional accompany of nucleolar proteins with NOL-58 26
2.	Direct evidence of ablated NOL-58 induces phenomena 27
3.	Potential mechanism of how nol-58 regulates the immune pathway 27
4.	The association of HLH-30 and NOL-58 28
5.	Box C/D RNP also regulated mitochondrial surveillance 29
6.	Future research in human cell lines 30
References 31
Figures 39
Figure 1. Flow chart of the nucleolar protein RNAi-based screening in C. elegans.  39
Figure 2. Decreased expression of most nucleolar proteins grants resistance to A. dhakensis infections in C. elegans.  41
Figure 3. The appearance of C. elegans under the microscope.  42
Figure 4. The measurement of physiologic characteristics shows differences between nol-58 silencing and wild-type nematodes.  44
Figure 5. The resistance to bacterial infections is not limited in A. dhakensis. 45
Figure 6. Knockdown nol-58 in tissue-specific strains and their resistance.	47
Figure 7. The immune pathways that were not associated with nol-58 in C. elegans against A. dhakensis infection. 48
Figure 8. Immune pathways that are epistatic to nol-58 in C. elegans with A. dhakensis infection. 50
Figure 9. The p38 phosphorylation is enhanced in nol-58 knockdown worms. 52
Figure 10. Relative mRNA expression of pmk-1 (p38 MAPK) downstream antimicrobial genes. 54
Figure 11. The FIB-1::GFP strain (cguIs001) and its survival. 56
Figure 12. The Nucleolar Surveillance Response (NSR) is reverted by nol-58 inhibition during A. dhakensis infection. 58
Figure 13. The silencing of nol-58 regresses the ability of Nucleolar Surveillance responses. 60
Figure 14. nol-58 is the most effective nucleolar protein on the list in terms of resistance to A. dhakensis. 61
Figure 15. The hlh-30 is not related to nol-58 in C. elegans. 63
Figure 16. Model of nol-58 regulates nucleolar surveillance response and p38 MAPK pathway in bacterial infection. 64
Tables 65
Table 1. The survival result of nucleolar protein screening in C. elegans. 65
Table 2. The survival result of nol-58 knockdown against multiple bacteria in C. elegans. 66
Table 3. The survival result of tissue-specific nol-58 knockdown in C. elegans. 67
Table 4. The survival result of nol-58 related epistasis analysis in C. elegans. 68
                                    

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