跳到主要內容

臺灣博碩士論文加值系統

(3.236.28.137) 您好!臺灣時間:2021/07/25 19:54
字體大小: 字級放大   字級縮小   預設字形  
回查詢結果 :::

詳目顯示

我願授權國圖
: 
twitterline
研究生:張翎君
研究生(外文):Lin-Chun Chang
論文名稱:MAGUK家族新成員4A其分子特性之研究
論文名稱(外文):Molecular characterization of 4A, a novel member of the MAGUK family
指導教授:林蔚靖
指導教授(外文):Wey-Jinq Lin
學位類別:碩士
校院名稱:國立陽明大學
系所名稱:生物藥學研究所
學門:生命科學學門
學類:生物科技學類
論文種類:學術論文
論文出版年:2004
畢業學年度:92
語文別:中文
中文關鍵詞:MAGUK
外文關鍵詞:MAGUK
相關次數:
  • 被引用被引用:0
  • 點閱點閱:88
  • 評分評分:
  • 下載下載:0
  • 收藏至我的研究室書目清單書目收藏:0
MAGUK家族的成員參與多樣的細胞過程,包括腫瘤抑制以及與不同訊息蛋白形成複合體以穩定細胞結構。由先前實驗室研究結果證實,以酵母菌雙雜合分析所選殖到會與抗增殖基因TIS21有交互作用的4A,是MAGUK家族的新成員。4A與另一個MAGUK家族成員MPP1有71%的胺基酸相同度。酵母菌雙雜合分析發現,hMPP1無法與TIS21產生交互作用。細胞體內過量表現MPP1蛋白,並不能如4A一樣具有使TIS21蛋白累積的作用。以專一性的引子對和反轉錄聚合酵素連鎖反應分析各種細胞株中4A及MPP1 mRNA的表現量,發現4A可在所有分析的細胞株中表現,而MPP1則除了小鼠及大鼠的fibroblast不被偵測到以外在其它細胞株也都看到表現。本研究合成hMPP1 peptide免疫兔子,利用純化的GST-hMPP1融合蛋白當作抗原進行抗體專一性之測試,在西方墨點標定法確認anti-hMPP1 peptide抗體可專一性辨認MPP1,而不會辨認到4A。
抗增殖基因TIS21可經由生長因子或分裂劑刺激而被迅速誘導。以蛋白質合成抑制劑cycloheximide處理,可見TIS21蛋白在NIH3T3細胞中的半衰期約為三十分鐘。根據先前實驗室研究的結果,在細胞中大量表現4A,可造成TIS21蛋白累積。因此以處理cycloheximide並同時表現4A蛋白,發現TIS21蛋白半衰期為大約六小時,顯見4A影響TIS21蛋白的穩定性,而促使TIS21的半衰期延長。利用proteasome抑制劑處理,確認TIS21的降解路徑經由ubiquitin-dependent pathway;而4A可能影響了TIS21蛋白被ubiquitination的步驟,使得TIS21的穩定性上升。而無法被CDK1磷酸化的TIS21S147A蛋白,同樣可被4A蛋白所穩定而延長其半衰期。大量表現4A蛋白於穩定表現TIS21蛋白的H1299細胞中,使細胞生長抑制更明顯。另外,4A可能也在細胞貼附的過程扮演一個角色。
Abstract
MAGUKs (membrane-associated guanylate kinase homologues) family members function in a variety of cellular processes including tumor suppression and receptor clustering. The previous results from ours lab reported that the rat 4A gene, cloned by a yeast two-hybrid screening using the antiproliferative gene TIS21 as a bait, encodes a novel member of MAGUK family. The deduced protein sequence of 4A shows 71% identity to that of mouse p55/MPP1. MPP1 does not interact with TIS21 in the yeast two-hybrid analysis. In contrast to 4A, MPP1 does not enhance the accumulation of TIS21 protein in vivo. The 4A and MPP1 messages are expressed at different levels in various human cell lines. Antibodies obtained using a peptide specific to hMPP1 recognizes MPP1 but not 4A in western blotting analysis using GST-hMPP1 as antigens.
TIS21 is an immediated-early gene with antiproliferative acitivity. In the presence of the protein synthesis inhibitor cycloheximide, the calculated half-life of TIS21 protein is 30 minutes in NIH3T3 cells. 4A has been shown to enhance accumulation of TIS21 proteins. In this studies, we show that 4A stabilizes TIS21 and prolongs its half-life to 6 hours. TIS21 proteins are degraded by ubiquitin-proteasome system in NIH3T3 cells. The TIS21S147A protein in which CDK1 phosphorylation site is mutated, shows a similar protein degradation pattern upon 4A overexpression. This indicates that the CDK1 phosphorylation site may not be involved in TIS21 degradation processes.
1. Lim, R. W., Varnum, B. C.,and Herschman, H. R. (1987) Cloning of tetradecanoyl phorbol ester-induced primary response sequence and their expression in density-arrested Swiss 3T3 cell and a TPA nonproliferative variant. Oncogene 1: 263-270
2. Kujubu, D. A., Lim, R. W., Varnum, B. C.,and Herschman, H. R. (1987) Induction of transiently expressed genes in PC-12 phenochromocytoma cells. Oncogene 1: 257-262
3. Herschman, H. R. (1991) Primary response genes induced by growth factors and tumor promoters. Ann. Rev. Biochem. 60: 281-319
4. Flecher, B. S., Lim, R. W., Varnum, B. C., Kujubu, D. A., Koski, R. A., and Herschman, H. R. (1991) Structure and expression of TIS21, a primary response gene induced by growth factors and tumor promoter. J. Biol. Chem. 266: 14511-14518
5. Varnum, B. C., Reddy, S. T., Koski, R. A., and Herschman, H. R. (1994) Synthesis, degradation, and subcellular localization of proteins encoded by the primary response genes TIS7/PC4 and TIS21/PC3. J. Cell. Physiol. 158: 205-213
6. Rouault, J. P., Falette, N., Guehenneux, F., Guillot, C., Rimokh, R., Wang, Q., Berthet, C., Moyret-Lalle, C., Savatier, P., Pain, B., Shaw, P., Berger, R., Samarut, J., Magaud, J. P., Ozturk, M., Samarut, C., and Puisieux, A. (1996) Identification of BTG2, an antiproliferative p53-dependent component of the DNA damage cellular response pathway. Nat. Genet. 14: 482-486
7. Montagnoli, A., Guardavaccaro, D., Starace, G., and Tirone, F. (1996) Overexpression of the nerve growth factor-inducible PC3 immediate early gene is assocaited with growth inhibition. Cell Growth Differ. 7: 1327-1336
8. Matsuda, S., Rouault, J., Magaud, J., and Berthet, C. (2001) In search of a function for the TIS21/PC3/BTG1/TOB family. FEBS Lett. 479: 67-72
9. Duriez, C., Falette, N., Audoynaud, C., Moyret-Lalle, C., Bensaad, K., Courtois, S., Wang,Q., Soussi, T., and Puisieux, A. (2002) The human BTG2/TIS21/PC3 gene : genomic structure, transcriptional regulation and evolution as a candidate tumor suppressor gene. Gene 282: 207-214
10. Lin, W.-J., Gary, J. D., Yang, M. C., Clarke, S., and Herschman, H. R. (1996) The mammalian immediate-early TIS21 protein and the leukemia-associated BTG1 protein interact with a protein-arginine N-methyltransferase. J. Biol. Chem. 271: 15034-15044
11. Lin, W.-J., Chang, Y.-F., Wang, W.-L., and Huang, C.-Y. F. (2001) Mitogen-stimulated TIS21 protein interacts with a protein-kinase-Cα-binding protein rPICK1. Biochem. J. 354: 635-643
12. Prevot,D.,Voeltzel, T., Birot, A.-M., Morel, A.-P., Rostan, M.-C., Magaud, J.-P., and Corbo, L. (2000) The leukemia-associated protein BTG1 and the p53-regulated protein BTG2 interacts with the homeoprotein Hoxb9 and enhance itstranscriptional activation. J. Biol. Chem. 275: 147-153
13. Rouault, J.-P., Prevot, D., Berhet, C., Birot, A.-M., Billaud, M., Magaud, J.-P., and Corbo, L. (1998) Interation of BTG1 and p53-regulated BTG2 gene products with mCaf1, the murine homolog of a component of the yeast CCR4 transcriptional regulatory complex. J. Biol. Chem.273: 22563-22569
14. Berthet, C., Guehenneux, F., Revol, V., samarut, C., Lukaszewicz, A., Dehay, C., Dumontet, C., Magaud, J.-P., and Mouault, J.-P. (2002) Interaction of PRMT1 with BTG/TOB proteins in cell signaling: molecular analysis and functional aspects. Genes to Cells. 7: 29-39
15. Cimato, T. R., Ettinger, M. J., Zhou, X., and Aletta, J. M. (1997) Nerve growth factor-specific regulation of protein methylation during neuronal differentiation of PC12 cells. J. Cell. Biol. 138: 1089-1103
16. Cimato, T. R., Tang, J., Xu, Y., Guarnaccia, C., Herschman, H. R., Pongor, S., and Aletta, J. M. (2002) Nerve growth factor-mediated increases in protein methylation occur predominantly at type I arginine methylation sites and involve protein arginine methyltransferase 1. J. Neurosci. Res. 67: 435-442
17. Staudinger, J., Zhou, J., Burgess, R., and Elledge, S. T. (1997) Specific interaction of the PDZ domain protein PICK1 with the COOH terminus of protein kinase C-α. J. Biol. Chem. 272: 32019-32024
18. Altin, J. G., Kujubu, D. A., Raffioni, S., Eveleth, D. D., Herschman, H. R., and Bradshaw, R. A. (1991) Differential induction of primary-response (TIS) gene in PC12 pheochromocytoma cells and the unresponsive variant PC12nnr5. J. Biol. Chem. 266: 5401-5406
19. Fanning, A. S., and Anderson, J. M. (1999) Protein modules as organizers of membrane structure. Curr. Opin. Cell Biol. 11: 432-439
20. Kuhlendahl, S., Spangenberg, O., Konrad, M., Kim, E., and Garner, C. C. (1998) Functional analysis of the guanylate-like domain in the synapse-associated protein SAP97. Eur. J. Biochem. 252: 305-313
21.Kistner, U., Garner, C. C., and Linial, M. (1995) Nucleotide binding by the synapse associated protein SAP90. FEBS Lett. 359: 159-163
22. Dimitratos, S. D., Woods, D. F., Stathakis, D. G., And Bryant, P. J. (1999) Signaling pathway are focused at specialized regions of the plasma membrane by scaffolding proteins of the MAGUK family. BioEssays 21: 912-921
23. Anderson, J.-M. (1996) Cell signalling: MAGUK magic. Curr. Biol. 6: 382-384
24. Sonyang, Z., Fanning, A. S., Fu, C., Xu, J., Marfatia, S. M., Chishiti, A. H., Crompton, A., Chan, A. C., Anderson, J. M., and Cantley, L. C. (1997) Recognition of unique carboxyl-terminal motifs by distinct PDZ domains. Science 275: 73-77
25. Pawson, T. (1994) SH2 and SH3 doamins in sginal transduction. Adv. Cancer Res. 64: 87-110
26. Hanada, T., Lin, L., Tibaldi, E. V., Reinherz, E. L., and Chishti, A. H. (2000) GAKIN: a novel kinesin-like protein associates with the human homologue of the Drosophila Disc Large tumor suppressor in T lyphocytes. J. Biol. Chem. 275: 28774-28784
27. Cohen, A. R., Woods, D. F., Marfatia, S. M., Walther, Z., Chishti, A. H., Anderson, J. M., and Wood, D. F. (1998) Human CASK/Lin-2 binds syndecan-2 and protein 4.1 and localizes to the basolateral membrane of epithelial cells. J. Cell Biol. 142: 129-138
28. Ruff, P., Speicher, D. W., and Chishti, A. H. (1991) Molecular identification of a major palmitoylated erythrocyte membrane protein contaning the src homology 3 motif. (1991) Proc. Natl. Acad. Sci. USA 88: 6595-6599
29. Kim, A. C., Metzenberg, A. B., Sahr, K. E., Marfatia, S. M., and Chishti, A. H. (1996) Complete genomic organization of the human erthroid p55 gene (MPP1), a membrane-associated guanylate kinase homologue. Genomics 31: 223-229
30. Alloisio, N., Venezia, N. D., Rana, A., Andrabi, K., Texier, P., Gilsanz, F., Cartron, J.-P., Delaunay, J., and Chishti, A. H. (1993) Evidence that the red blood cell protein p55 may participate in the skeleton-membrane linkage that involves protein 4.1 and glycophorin C. Blood 82: 1323-1327
31. Marfatia, S. M., Lue, R. A., Branton, D., and Chishti, A. H. (1994) In vitro binding studies suggest a membrane-associated complex betweenerythroid p55, protein 4.1, and glycophorin C. J. Biol. Chem. 269: 8631-8634
32. Marfatia, S. M., Lue, R. A., Branton, D., and Chishti, A. H. (1995) Identification of the protein 4.1 binding interafce on glycophorin C and p55, a homologue of the Drosophila disc-large tumor suppressor protein. J. Biol. Chem. 270: 715-719
33. Nunomura, W., Takakuwa, Y., Parra, M., Conboy, J., and Mohandas, N. (2000) Regulation of protein 4.1R, p55, and glycophorin C ternary complex in human erythrocyte membrane. J. Biol. Chem. 275: 24540-24546
34. Marfatia, S. M., Morais-Cabral J. H., Kim, A. C., and Byron, O. (1997) The PDZ domain of human erythrocyte p55 mediates its binding to the cytoplasmic carboxyl terminus of glycophorin C. J. Biol. Chem. 272: 24191-24197
35. Woods, D. F., and Bryant, P. J. (1991) The disc-large tumor suppressor gene of Drosophila encodes a guanylate kinase homolog localized at septate junction. Cell 66: 451-464
36. Kaech, S. M., Whitfield, C. W., and Kim, S. K. (1998) Basolateral membranelocalization of the receptor tyrosine kinase Let-23 is mediated by the Lin-2/Lin-7/Lin-10 protein complex in the vulval precursor cells of C. elegans. Cell 94: 761-771
37. Hsueh, Y. P., Wang, T. F., Yang, F. C., and Sheng, M. (2000) Nuclear translocation and transcription regulation by the membrane-associated guanylate kinase CASK/Lin-2. Nature 404: 298-302
38. Lue, R. A., Marfatia, S. M., Branton, D., and Chishti, A. H. (1994) Cloning and Characterization of hdlg: the human homologue of the Drosophila disc large tumor suppressor binds to protein 4.1 Proc. Natl. Acad. Sci. USA 91: 9818-9822
39. Pickart, C. M. (2001) Mechanisms underlying ubiquitination. Ann Rev. Biochem. 70: 503-533
40. Hochstrasser, M. (1995) Ubiquitin-dependent protein degradation. Ann. Rev.Genet. 30: 405-439
41. Ficazzola, M. A., Fraiman, M., Gitlin, J., Woo, K., Melamed, J., Rubin, M. A., and Walden, P. D. (2001) Antiproliferative B cell translocation gene 2 protein is down-regulated post-transcriptionally as an early event in prostate carcinogenesis. Carcinogenesis 22: 1271-1279
42. Hirokawa, N., Noda, Y., and Okada, Y. (1998) Kinesin and dynein superfamily proteins in organelle transport and cell division. Curr. Opin. Cell Biol. 10: 60-73
43. Elder, B., Kuo, K., Gitschier, J., Kim, A., Chishti, A., and Metzenberg, A. (1996) cDNA sequence and genomic structure of the murine p55 (Mpp1) gene. Genomics 38: 231-234
44. Sasajima, H., Nakagawa, K., and Yokosawa, H. (2002) Antiproliferative proteins of the BTG/Tob family are degraded by the ubiquitin-proteasome system. Eur J Biochem. 269: 3596-604
45. Gaide O, Martinon F, Micheau O, Bonnet D, Thome M, Tschopp J. (2001) Carma1, a CARD-containing binding partner of Bcl10, induces Bcl10 phosphorylation and NF-κB activation FEBS Lett. 496, 121-127.
46. Bertin J, Wang L, Guo Y, Jacobson MD, Poyet JL, Srinivasula SM, Merriam S, DiStefano PS, Alnemri (2001) CARD11 and CARD14 are novel caspase recruitment domain (CARD)/membrane-associated guanylate kinase (MAGUK) family members that interact with Bcl10 and activate NF-κB. J Biol Chem. 276: 11877-11882.
47. McAllister-Lucas LM, Inohara N, Lucas PC, Ruland J, Benito A, Li Q, Chen S,Chen FF, Yamaoka S, Verma IM, Mak TW, Nunez G. (2001) Bimp1, a MAGUK family member linking protein kinase C activation to Bcl10-mediated NF-κB induction. J Biol Chem. 276: 30589-30597.
48. Wang L, Guo Y, Huang WJ, Ke X, Poyet JL, Manji GA, Merriam S, Glucksmann MA, DiStefano PS, Alnemri ES, Bertin J. (2001) CARD10 is a novel caspase recruitment domain/membrane-associated guanylate kinase family member that interacts with BCL10 and activates NF-kappa B. J Biol Chem. 276: 21405-21409.
49.Stilo R, Liguoro D, Di Jeso B, Formisano S, Consiglio E, Leonardi A, Vito P. (2004) Physical and functional interaction of CARMA1 and CARMA3 with IKKgamma -NEMO. J Biol Chem. (in pressed)
50. McGee, A. W., and Bredt, D. S. (1999) Identification of an intracellular interaction between the SH3 and guanylate kinase domains of PSD-95. J Biol Chem. 274: 17431-17436
51. Nix, S. L., Chishti, A. H. Anderson, J. M. and Walther, Z. (2000) hCASK and hDlg associate in epithelia, and their src homology 3 and guanylate kinase domains participate in both intracellular and intermolecular interactions. J Biol Chem. 275: 41192-41200
52. Shin, H., Hsueh, Y.-P., Yang, F.-C. Kim, E. and Sheng, M. (2000) An intramolecular interaction between src homology 3 domain and guanylate kinase-like domain required for channel clustering by postsynaptic density -95/SAP90. J. Neurosci. 20: 3580-3587
53. McGee, A. W., Dakoji, S. R., Olsen, O., Bredt, D. S., Lim, W. A., and Prehoda, K. E. (2001) Structure of the SH3-guanylate kinase module from PSD-95 suggests a mechanism for regulated assembly of MAGUK scaffolding proteins. Mol. Cell 8: 1291-1301
54. Wu, H., Reissner, C., Kuhlendahl, S., Coblentz, B., Reuver, S., Kindler, S., Gundelfinger, E. D., and Garner, C. C. (2000) Intramolecular interactions regulate SAP97 binding to GKAP. EMBO J. 19: 5740-5751
55. Wu, H., Reuver, S. M., Kuhlendahl, S., Chung, W. J., and Garner, C. C. (1998) Subcellular targeting and cytoskeletal attachment of SAP97 to the epithelial lateral membrane. J.Cell Sci. 111: 2365-2376
56. El-Husseini, A. E., Topinka, J. R., Lehrer-Graiwer, J. E., Firestein, B. L., Craven, S. E., Aoki, C. and Bredt, D. S. (2000) Ion channel clustering by membrane -associated guanylate kinases. J Biol Chem. 275: 23904–23910
57. 梁淑菁 (1999) TIS21 及 BTG1 基因表現與鄰酸化之研究 碩士論文 台北 : 國立陽明大學生物藥學所 36-41
58. 蕭舜方 (2003) TIS21基因對於H1299細胞生長以及藥物誘導細胞死亡的影響 碩士論文 台北 : 國立陽明大學生物藥學所 24-31
59. 王春發 (2002) MAGUK家族新成員的鑑定及其與早期立即基因TIS21交互作用之研究 碩士論文 台北 : 國立陽明大學生物藥學所 34-45
60. Johnsson, N. and Varshavsky, A. (1994) Split ubiquitin as a sensor of protein interactions in vivo. Proc. Natl. Acad. Sci. USA 91: 10340-10344
61. Stagljar, I., Korostensky, C., Johnsson, N. and Heesen, S. (1998) A genetic systerm based on split-ubiquitin for the analysis of interactions between membrane proteins in vivo. Proc. Natl. Acad. Sci. USA 95: 5187-5192
62. Kohler, F. and Muller, K. M. (2003) Adaption of the Ras-recruitment system to the analsyis of interactions between membrane-associated proteins. Nucleic Acids Res. 31: e28
63. Stagljar, I. and Fields, S. (2002) Analysis of membrane protein interactions using yeast-based technologies. Trends in Biochem. Sci. 27: 559-563
QRCODE
 
 
 
 
 
                                                                                                                                                                                                                                                                                                                                                                                                               
第一頁 上一頁 下一頁 最後一頁 top