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| ???org.dspace.app.webui.jsptag.ItemTag.dcfield??? | Value | Language |
|---|---|---|
| dc.contributor.advisor | 李芳仁 | |
| dc.contributor.author | Yi-Yun Liu | en |
| dc.contributor.author | 劉宜昀 | zh_TW |
| dc.date.accessioned | 2021-06-08T04:51:27Z | - |
| dc.date.copyright | 2009-09-15 | |
| dc.date.issued | 2009 | |
| dc.date.submitted | 2009-07-27 | |
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| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/23281 | - |
| dc.description.abstract | 真核細胞中，基因的調控需要核醣核酸結合蛋白與核醣核酸形成複合體，稱為核醣核蛋白。核醣核酸結合蛋白(Rbp1p)轉譯成一個六百七十二個胺基酸組成，約八十kD的蛋白質；包含三個核醣核酸識別基序 (RRM)及兩個富含麩氨酸區域。核醣核酸結合蛋白(Rbp1p)在發現時，被定義為一個負調控生長的因子。在我們實驗室中，已經發現在細胞內大量表現核醣核酸結合蛋白(Rbp1p) 能夠抑制酵母菌生長、促進粒腺體外模孔蛋白(porin)訊息核醣核酸降解、並抑制蛋白質的轉譯。核醣核酸結合蛋白(Rbp1p)在細胞中會座落到細胞質中特殊的聚集顆粒，稱為P-bodies。本實驗中，我們發現在第四百二十八個胺基酸由絲胺酸變成脯胺酸的突變株，會部份或完全喪失上述已知三項核醣核酸結合蛋白(Rbp1p)的功能。但這樣的核醣核酸結合蛋白(Rbp1p)突變株依然能夠座落到P-bodies。我們也發現：核醣核酸結合蛋白(Rbp1p)的絲胺酸-脯胺酸突變株及三個核醣核酸識別基序(RRM) 突變株均能夠引發絮凝現象(flocculation) 及洋菜入侵生長現象(invasive growth)這兩種附著現象。絮凝基因(Flo1p)和(Flo11p)是細胞膜上能夠進行附著的糖化蛋白。絮凝基因(Flo1p)剔除細胞株表現核醣核酸結合蛋白絲胺酸-脯胺酸突變株時不能夠引發絮凝現象；絮凝基因(Flo11p)剔除細胞株表現核醣核酸結合蛋白絲胺酸-脯胺酸突變株時則會降低洋菜入侵生長現象。絮凝基因(Flo1p)和(Flo11p)共同的轉錄因子(Mss11p)對於絲胺酸-脯胺酸突變株引發的附著現象也是必需的，推測核醣核酸結合蛋白(Rbp1p)的絲胺酸-脯胺酸突變株透過調控轉錄因子MSS11引發絮凝現象及洋菜入侵生長現象。與核醣核酸結合蛋白(Rbp1p)有交互作用的蛋白質中，Dhh1p、Xrn1p和Kre6p是可能的目標蛋白，藉由與絲胺酸-脯胺酸突變株及三個核醣核酸識別基序(RRM) 突變株的交互作用，調控附著現象。 | zh_TW |
| dc.description.abstract | RNA-binding proteins forming dynamic messenger ribonucleoproteins with the transcript are required for the regulation of eukaryotic gene expression. The RNA-binding protein RBP1 encodes a 672-amino acid, ~80-kD protein containing three RNA recognition motifs (RRM) and two glutamine-rich stretches. It is first identified as a negative growth regulator. Our lab has demonstrated overexpression of Rbp1p shows a slow-growth phenotype, decreases the porin mRNA level by enhancing degradation, and suppresses the translation. Rbp1p localized to cytoplasm foci, named P-bodies. Here we showed that serine-428-Proline mutation loses the function of Rbp1p in slow-growth, translation repression and decreasing the stability of POR1 mRNA, but it still can localize to P-bodies. Rbp1p-S428P and Rbp1p RRM motif mutants induce flocculation and invasive growth. Flocculins, FLO1 and FLO11, are membrane surface glycoproteins for adhesion. FLO1 deletion suppresses flocculation; however FLO11 deletion decreases the invasive-growth level. It is also demonstrated that MSS11, transcription factor of FLO1 and FLO11 is required for both adhesion phenotypes, suggesting RNA-binding proteins mutation might regulate flocculation and invasive growth through MSS11. Rbp1p interacting proteins, DHH1, XRN1 and KRE6 might be possible candidates interacting with Rbp1p-S428P and Rbp1p RRM motif mutants to regulate adhesion phenotypes. | en |
| dc.description.provenance | Made available in DSpace on 2021-06-08T04:51:27Z (GMT). No. of bitstreams: 1 ntu-98-R96448003-1.pdf: 3102748 bytes, checksum: d5e25dd72318e567e6184dd349156bfa (MD5) Previous issue date: 2009 | en |
| dc.description.tableofcontents | Table of Contents .....................................II
中文摘要 ...............................................IV Abstract ................................................V Introduction.............................................1 Materials and Methods....................................9 Results.................................................17 Discussion .............................................26 Figures.................................................31 Figure 1. Rbp1p-S428P shows no growth inhibition in YPH499. ................................................31 Figure 2. Serine-428-Proline mutation partially loses the functions of Rbp1p. ....................................32 Figure 3. P-bodies localization of Rbp1p is not affected by mutation on Ser-428-Proline. ........................33 Figure 4. Rbp1p-S428P induces flocculation and invasive growth in BY4741. ......................................34 Figure 5. Rbp1p-S428P induces flocculation and invasive growth in YPH499. ......................................35 Figure 6. Rbp1p-RRMs mutations also induce flocculation and invasive growth. ...................................36 Figure 7. FLO1 is required for RBP1-S428P induced flocculation; FLO11 is required for RBP1-S428P induced invasive growth in BY4741. .............................37 Figure 8. MSS11 is required for RBP1-S428P induced flocculation and invasive growth in BY4741. ...........39 Figure 9. FLO1 and MSS11 are required for RBP1-S428P induced flocculation and invasive growth in YPH499.......40 Figure 10. Steady-state mRNA level of indicated strains expressing HA-Rbp1p and -S428P. .........................41 Figure 11. TPK2, TPK3, KSS1, and SFL1 could regulate RBP1-S428P induced flocculation, but not invasive growth. ....42 Figure 12. DHH1 and KRE6 are required for RBP1-S428P induced flocculation and invasive growth in BY4741. .....43 Figure 13. DHH1 and KRE6 are required for RBP1-rrm1, -rrm2 and -rrm3 induced flocculation and invasive growth in BY4741. ................................................44 Figure 14. Rbp1p-SPSA decreases the interaction with Rbp1p interacting proteins, Dhh1p and Kre6p. ..................45 Figure 15. XRN1 is required for RBP1-S428P induced flocculation and invasive growth in BY4741. .............46 Figure 16. XRN1 is required for RBP1-rrm1, -rrm2 and -rrm3 induced flocculation and invasive growth in BY4741. .....47 Figure 17. DHH1 and XRN1 are required for RBP1-S428P induced flocculation and invasive growth in YPH499. .....48 Figure 18. Models of this study ....................49 Appendix 1. Overexpression of HA-RBP1 suppresses Σ1278b invasive growth and decreases FLO11 RNA level. ..........50 Appendix 2. Rbp1p-S428P and –rrm2 precipitates less Dhh1p than Rbp1p full-length in immunoprecipitation. ..........51 Appendix 3. P-bodies localization of Rbp1p is not affected by mutation on Ser-428-Proline. .........................52 Table 1. Yeast strains used in this study ...............53 Table 2. Primers used in this study .....................54 Table 3. A brief summary of plasmids used in this study..56 Table 4. Antibodies used in this study...................57 References...............................................58 | |
| dc.language.iso | en | |
| dc.subject | 核醣核酸結合蛋白 | zh_TW |
| dc.subject | 絮凝現象 | zh_TW |
| dc.subject | 核醣核酸識別基序 | zh_TW |
| dc.subject | 洋菜入侵生長現象 | zh_TW |
| dc.subject | invasive growth | en |
| dc.subject | RNA-binding proteins | en |
| dc.subject | RNA recognition motifs | en |
| dc.subject | flocculation | en |
| dc.title | 酵母菌核醣核酸蛋白Rbp1p絲胺酸-脯胺酸及核醣核酸辨識區域突變株之功能探討 | zh_TW |
| dc.title | Functional characterization of Ser-428-Pro and RRMs mutations in Rbp1p | en |
| dc.type | Thesis | |
| dc.date.schoolyear | 97-2 | |
| dc.description.degree | 碩士 | |
| dc.contributor.oralexamcommittee | 鄧述諄,張典顯,鄭明媛,羅?升 | |
| dc.subject.keyword | 核醣核酸結合蛋白,核醣核酸識別基序,絮凝現象,洋菜入侵生長現象, | zh_TW |
| dc.subject.keyword | RNA-binding proteins,RNA recognition motifs,flocculation,invasive growth, | en |
| dc.relation.page | 63 | |
| dc.rights.note | 未授權 | |
| dc.date.accepted | 2009-07-27 | |
| dc.contributor.author-college | 醫學院 | zh_TW |
| dc.contributor.author-dept | 分子醫學研究所 | zh_TW |
| Appears in Collections: | 分子醫學研究所 | |
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