請用此 Handle URI 來引用此文件:
http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/16453
完整後設資料紀錄
DC 欄位 | 值 | 語言 |
---|---|---|
dc.contributor.advisor | 周子賓(Tze-Bin Chou) | |
dc.contributor.author | Lin-Hsiang Chuang | en |
dc.contributor.author | 莊淋翔 | zh_TW |
dc.date.accessioned | 2021-06-07T18:15:37Z | - |
dc.date.copyright | 2012-03-19 | |
dc.date.issued | 2012 | |
dc.date.submitted | 2012-02-17 | |
dc.identifier.citation | References
Albertson, R., Riggs, B., & Sullivan, W. (2005). Membrane traffic: a driving force in cytokinesis. [Review]. Trends Cell Biol, 15(2), 92-101. Andrews, H. K., Zhang, Y. Q., Trotta, N., & Broadie, K. (2002). Drosophila sec10 is required for hormone secretion but not general exocytosis or neurotransmission. Traffic, 3(12), 906-921. Arlaud, G. J., Volanakis, J. E., Thielens, N. M., Narayana, S. V., Rossi, V., & Xu, Y. (1998). The atypical serine proteases of the complement system. Adv Immunol, 69, 249-307. Barr, F. A., & Gruneberg, U. (2007). Cytokinesis: placing and making the final cut. [Research Support, Non-U.S. Gov'tReview]. Cell, 131(5), 847-860. Bell, A. W., Ward, M. A., Blackstock, W. P., Freeman, H. N., Choudhary, J. S., Lewis, A. P., . . . Bergeron, J. J. (2001). Proteomics characterization of abundant Golgi membrane proteins. [Research Support, Non-U.S. Gov't]. J Biol Chem, 276(7), 5152-5165. Beronja, S., Laprise, P., Papoulas, O., Pellikka, M., Sisson, J., & Tepass, U. (2005). Essential function of Drosophila Sec6 in apical exocytosis of epithelial photoreceptor cells. J Cell Biol, 169(4), 635-646. Bonangelino, C. J., Chavez, E. M., & Bonifacino, J. S. (2002). Genomic screen for vacuolar protein sorting genes in Saccharomyces cerevisiae. [Research Support, Non-U.S. Gov't]. Mol Biol Cell, 13(7), 2486-2501. Bornemann, D. J., Duncan, J. E., Staatz, W., Selleck, S., & Warrior, R. (2004). Abrogation of heparan sulfate synthesis in Drosophila disrupts the Wingless, Hedgehog and Decapentaplegic signaling pathways. [Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.]. Development, 131(9), 1927-1938. Boyd, C., Hughes, T., Pypaert, M., & Novick, P. (2004). Vesicles carry most exocyst subunits to exocytic sites marked by the remaining two subunits, Sec3p and Exo70p. J Cell Biol, 167(5), 889-901. Brand, A. H., & Perrimon, N. (1993). Targeted gene expression as a means of altering cell fates and generating dominant phenotypes. Development, 118(2), 401-415. Burgess, R. W., Deitcher, D. L., & Schwarz, T. L. (1997). The synaptic protein syntaxin1 is required for cellularization of Drosophila embryos. J Cell Biol, 138(4), 861-875. Chang, A. J., & Morisato, D. (2002). Regulation of Easter activity is required for shaping the Dorsal gradient in the Drosophila embryo. Development, 129(24), 5635-5645. Chang, C. W. (2007). Drosophila Golgi protein, Rotini, regulates glycosaminoglycan chains polymerases in the biogenesis of Heparan sulfate proteoglycans. Master Thesis. National Taiwan University, Taiwan. Chang, S. C. (2011). The approach of Drosophila GOLPH3, Rotini, in modulating Dorsal-Ventral axis formation. Master Thesis. National Taiwan University, Taiwan. Chasan, R., & Anderson, K. V. (1989). The role of easter, an apparent serine protease, in organizing the dorsal-ventral pattern of the Drosophila embryo. Cell, 56(3), 391-400. Chou, T. B., Noll, E., & Perrimon, N. (1993). Autosomal P[ovoD1] dominant female-sterile insertions in Drosophila and their use in generating germ-line chimeras. Development, 119(4), 1359-1369. Chou, T. B., & Perrimon, N. (1996). The autosomal FLP-DFS technique for generating germline mosaics in Drosophila melanogaster. Genetics, 144(4), 1673-1679. Cockcroft, S., Taylor, J. A., & Judah, J. D. (1985). Subcellular localisation of inositol lipid kinases in rat liver. Biochim Biophys Acta, 845(2), 163-170. D'Angelo, G., Vicinanza, M., Di Campli, A., & De Matteis, M. A. (2008). The multiple roles of PtdIns(4)P -- not just the precursor of PtdIns(4,5)P2. J Cell Sci, 121(Pt 12), 1955-1963. De Matteis, M. A., & Godi, A. (2004). PI-loting membrane traffic. Nat Cell Biol, 6(6), 487-492. DeLotto, R. (2001). Gastrulation defective, a complement factor C2/B-like protease, interprets a ventral prepattern in Drosophila. EMBO Rep, 2(8), 721-726. DeLotto, R., & Spierer, P. (1986). A gene required for the specification of dorsal-ventral pattern in Drosophila appears to encode a serine protease. Nature, 323(6090), 688-692. Demmel, L., Gravert, M., Ercan, E., Habermann, B., Muller-Reichert, T., Kukhtina, V., . . . Walch-Solimena, C. (2008). The clathrin adaptor Gga2p is a phosphatidylinositol 4-phosphate effector at the Golgi exit. Mol Biol Cell, 19(5), 1991-2002. DiMario, P. J., & Mahowald, A. P. (1987). Female sterile (1) yolkless: a recessive female sterile mutation in Drosophila melanogaster with depressed numbers of coated pits and coated vesicles within the developing oocytes. J Cell Biol, 105(1), 199-206. Dippold, H. C., Ng, M. M., Farber-Katz, S. E., Lee, S. K., Kerr, M. L., Peterman, M. C., . . . Field, S. J. (2009). GOLPH3 bridges phosphatidylinositol-4- phosphate and actomyosin to stretch and shape the Golgi to promote budding. Cell, 139(2), 337-351. Dissing, M., Giordano, H., & DeLotto, R. (2001). Autoproteolysis and feedback in a protease cascade directing Drosophila dorsal-ventral cell fate. EMBO J, 20(10), 2387-2393. Finger, F. P., Hughes, T. E., & Novick, P. (1998). Sec3p is a spatial landmark for polarized secretion in budding yeast. Cell, 92(4), 559-571. Finger, F. P., & White, J. G. (2002). Fusion and fission: membrane trafficking in animal cytokinesis. [Review]. Cell, 108(6), 727-730. Foe, V. E., & Alberts, B. M. (1983). Studies of nuclear and cytoplasmic behaviour during the five mitotic cycles that precede gastrulation in Drosophila embryogenesis. J Cell Sci, 61, 31-70. Grindstaff, K. K., Yeaman, C., Anandasabapathy, N., Hsu, S. C., Rodriguez-Boulan, E., Scheller, R. H., & Nelson, W. J. (1998). Sec6/8 complex is recruited to cell-cell contacts and specifies transport vesicle delivery to the basal-lateral membrane in epithelial cells. Cell, 93(5), 731-740. Guo, W., & Novick, P. (2004). The exocyst meets the translocon: a regulatory circuit for secretion and protein synthesis? Trends Cell Biol, 14(2), 61-63. Guo, W., Roth, D., Walch-Solimena, C., & Novick, P. (1999). The exocyst is an effector for Sec4p, targeting secretory vesicles to sites of exocytosis. [Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.]. EMBO J, 18(4), 1071-1080. Guo, W., Tamanoi, F., & Novick, P. (2001). Spatial regulation of the exocyst complex by Rho1 GTPase. [Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.]. Nat Cell Biol, 3(4), 353-360. Hama, H., Schnieders, E. A., Thorner, J., Takemoto, J. Y., & DeWald, D. B. (1999). Direct involvement of phosphatidylinositol 4-phosphate in secretion in the yeast Saccharomyces cerevisiae. J Biol Chem, 274(48), 34294-34300. Hammond, G. R., Schiavo, G., & Irvine, R. F. (2009). Immunocytochemical techniques reveal multiple, distinct cellular pools of PtdIns4P and PtdIns(4,5)P(2). Biochem J, 422(1), 23-35. Han, J. H., Lee, S. H., Tan, Y. Q., LeMosy, E. K., & Hashimoto, C. (2000). Gastrulation defective is a serine protease involved in activating the receptor toll to polarize the Drosophila embryo. Proc Natl Acad Sci U S A, 97(16), 9093-9097. Hashimoto, C., Kim, D. R., Weiss, L. A., Miller, J. W., & Morisato, D. (2003). Spatial regulation of developmental signaling by a serpin. Dev Cell, 5(6), 945-950. Hsu, S. C., TerBush, D., Abraham, M., & Guo, W. (2004). The exocyst complex in polarized exocytosis. [Review]. Int Rev Cytol, 233, 243-265. Inoue, M., Chang, L., Hwang, J., Chiang, S. H., & Saltiel, A. R. (2003). The exocyst complex is required for targeting of Glut4 to the plasma membrane by insulin. Nature, 422(6932), 629-633. Konrad, K. D., Goralski, T. J., Mahowald, A. P., & Marsh, J. L. (1998). The gastrulation defective gene of Drosophila melanogaster is a member of the serine protease superfamily. Proc Natl Acad Sci U S A, 95(12), 6819-6824. Krem, M. M., & Di Cera, E. (2002). Evolution of enzyme cascades from embryonic development to blood coagulation. Trends Biochem Sci, 27(2), 67-74. Kristen, U., & Lockhausen, J. (1983). Estimation of Golgi membrane flow rates in ovary glands of aptenia cordifolia using cytochalasin B. Eur J Cell Biol, 29(2), 262-267. Langevin, J., Morgan, M. J., Sibarita, J. B., Aresta, S., Murthy, M., Schwarz, T., . . . Bellaiche, Y. (2005). Drosophila exocyst components Sec5, Sec6, and Sec15 regulate DE-Cadherin trafficking from recycling endosomes to the plasma membrane. Dev Cell, 9(3), 365-376. Lecuit, T., & Wieschaus, E. (2000). Polarized insertion of new membrane from a cytoplasmic reservoir during cleavage of the Drosophila embryo. [Research Support, Non-U.S. Gov't]. J Cell Biol, 150(4), 849-860. Lemmon, M. A. (2008). Membrane recognition by phospholipid-binding domains. Nat Rev Mol Cell Biol, 9(2), 99-111. LeMosy, E. K., Hong, C. C., & Hashimoto, C. (1999). Signal transduction by a protease cascade. [Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.Review]. Trends Cell Biol, 9(3), 102-107. Ligoxygakis, P., Roth, S., & Reichhart, J. M. (2003). A serpin regulates dorsal-ventral axis formation in the Drosophila embryo. Curr Biol, 13(23), 2097-2102. Lipschutz, J. H., Guo, W., O'Brien, L. E., Nguyen, Y. H., Novick, P., & Mostov, K. E. (2000). Exocyst is involved in cystogenesis and tubulogenesis and acts by modulating synthesis and delivery of basolateral plasma membrane and secretory proteins. Mol Biol Cell, 11(12), 4259-4275. Misra, S., Hecht, P., Maeda, R., & Anderson, K. V. (1998). Positive and negative regulation of Easter, a member of the serine protease family that controls dorsal-ventral patterning in the Drosophila embryo. Development, 125(7), 1261-1267. Moussian, B., & Roth, S. (2005). Dorsoventral axis formation in the Drosophila embryo--shaping and transducing a morphogen gradient. Curr Biol, 15(21), R887-899. Murthy, M., & Schwarz, T. L. (2004). The exocyst component Sec5 is required for membrane traffic and polarity in the Drosophila ovary. Development, 131(2), 377-388. Murthy, M., Teodoro, R. O., Miller, T. P., & Schwarz, T. L. (2010). Sec5, a member of the exocyst complex, mediates Drosophila embryo cellularization. [Research Support, N.I.H., Extramural]. Development, 137(16), 2773-2783. Neuman-Silberberg, F. S., & Schupbach, T. (1994).Dorsoventral axis formation in Drosophila depends on the correct dosage of the gene. Development, 120(9), 2457-2463. Nilson, L. A., & Schupbach, T. (1999). EGF receptor signaling in Drosophila oogenesis. [Review]. Curr Top Dev Biol, 44, 203-243. Novick, P., Field, C., & Schekman, R. (1980). Identification of 23 complementation groups required for post-translational events in the yeast secretory pathway. Cell, 21(1), 205-215. Pai, L. M., Barcelo, G., & Schupbach, T. (2000). D-cbl, a negative regulator of the Egfr pathway, is required for dorsoventral patterning in Drosophila oogenesis. Cell, 103(1), 51-61. Peri, F., Technau, M., & Roth, S. (2002). Mechanisms of Gurken-dependent pipe regulation and the robustness of dorsoventral patterning in Drosophila. Development, 129(12), 2965-2975. Perrimon, N., & Perkins, L. A. (1997). There must be 50 ways to rule the signal: the case of the Drosophila EGF receptor. Cell, 89(1), 13-16. Prigent, M., Dubois, T., Raposo, G., Derrien, V., Tenza, D., Rosse, C., . . . Chavrier, P. (2003). ARF6 controls post-endocytic recycling through its downstream exocyst complex effector. J Cell Biol, 163(5), 1111-1121. Queenan, A. M., Barcelo, G., Van Buskirk, C., & Schupbach, T. (1999). The transmembrane region of Gurken is not required for biological activity, but is necessary for transport to the oocyte membrane in Drosophila. Mech Dev, 89(1-2), 35-42. Rose, T., LeMosy, E. K., Cantwell, A. M., Banerjee-Roy, D., Skeath, J. B., & Di Cera, E. (2003). Three-dimensional models of proteases involved in patterning of the Drosophila Embryo. Crucial role of predicted cation binding sites. J Biol Chem, 278(13), 11320-11330. Roth, S., Neuman-Silberberg, F. S., Barcelo, G., & Schupbach, T. (1995). cornichon and the EGF receptor signaling process are necessary for both anterior-posterior and dorsal-ventral pattern formation in Drosophila. Cell, 81(6), 967-978. Roth, S., & Schupbach, T. (1994). The relationship between ovarian and embryonic dorsoventral patterning in Drosophila. Development, 120(8), 2245-2257. Roth, T. F., & Porter, K. R. (1964). YOLK PROTEIN UPTAKE IN THE OOCYTE OF THE MOSQUITO AEDES AEGYPTI. L. J Cell Biol, 20, 313-332. Ruohola-Baker, H., Grell, E., Chou, T. B., Baker, D., Jan, L. Y., & Jan, Y. N. (1993). Spatially localized rhomboid is required for establishment of the dorsal-ventral axis in Drosophila oogenesis. Cell, 73(5), 953-965. Salminen, A., & Novick, P. J. (1989). The Sec15 protein responds to the function of the GTP binding protein, Sec4, to control vesicular traffic in yeast. J Cell Biol, 109(3), 1023-1036. Santiago-Tirado, F. H., Legesse-Miller, A., Schott, D., & Bretscher, A. (2011). PI4P and Rab inputs collaborate in myosin-V-dependent transport of secretory compartments in yeast. Dev Cell, 20(1), 47-59. Schmitz, K. R., Liu, J., Li, S., Setty, T. G., Wood, C. S., Burd, C. G., & Ferguson, K. M. (2008). Golgi localization of glycosyltransferases requires a Vps74p oligomer. Dev Cell, 14(4), 523-534. Schonbaum, C. P., Perrino, J. J., & Mahowald, A. P. (2000). Regulation of the vitellogenin receptor during Drosophila melanogaster oogenesis. Mol Biol Cell, 11(2), 511-521. Schupbach, T. (1987). Germ line and soma cooperate during oogenesis to establish the dorsoventral pattern of egg shell and embryo in Drosophila melanogaster. Cell, 49(5), 699-707. Schweitzer, R., Howes, R., Smith, R., Shilo, B. Z., & Freeman, M. (1995). Inhibition of Drosophila EGF receptor activation by the secreted protein Argos. Nature, 376(6542), 699-702. Scott, K. L., & Chin, L. (2010). Signaling from the Golgi: mechanisms and models for Golgi phosphoprotein 3-mediated oncogenesis. Clin Cancer Res, 16(8), 2229-2234. Scott, K. L., Kabbarah, O., Liang, M. C., Ivanova, E., Anagnostou, V., Wu, J., . . . Chin, L. (2009). GOLPH3 modulates mTOR signalling and rapamycin sensitivity in cancer. Nature, 459(7250), 1085-1090. Sen, J., Goltz, J. S., Stevens, L., & Stein, D. (1998). Spatially restricted expression of pipe in the Drosophila egg chamber defines embryonic dorsal-ventral polarity. Cell, 95(4), 471-481. Smith, C., Giordano, H., & DeLotto, R. (1994). Mutational analysis of the Drosophila snake protease: an essential role for domains within the proenzyme polypeptide chain. Genetics, 136(4), 1355-1365. Smith, C. L., Giordano, H., Schwartz, M., & DeLotto, R. (1995). Spatial regulation of Drosophila snake protease activity in the generation of dorsal-ventral polarity. Development, 121(12), 4127-4135. Snyder, C. M., Mardones, G. A., Ladinsky, M. S., & Howell, K. E. (2006). GMx33 associates with the trans-Golgi matrix in a dynamic manner and sorts within tubules exiting the Golgi. [Research Support, N.I.H., Extramural]. Mol Biol Cell, 17(1), 511-524. Sommer, B., Oprins, A., Rabouille, C., & Munro, S. (2005). The exocyst component Sec5 is present on endocytic vesicles in the oocyte of Drosophila melanogaster. J Cell Biol, 169(6), 953-963. Song, H. H. (2001). A novel putative Golgi gene, rotini, is required for the production and transportation of Drosophila Hedgehog. Master Thesis. National Taiwan University, Taiwan. Stein, D., & Nusslein-Volhard, C. (1992). Multiple extracellular activities in Drosophila egg perivitelline fluid are required for establishment of embryonic dorsal-ventral polarity. Cell, 68(3), 429-440. Stroud, R. M., Kossiakoff, A. A., & Chambers, J. L. (1977). Mechanisms of zymogen activation. Annu Rev Biophys Bioeng, 6, 177-193. Szentpetery, Z., Varnai, P., & Balla, T. (2010). Acute manipulation of Golgi phosphoinositides to assess their importance in cellular trafficking and signaling. Proc Natl Acad Sci U S A, 107(18), 8225-8230. Takei, Y., Ozawa, Y., Sato, M., Watanabe, A., & Tabata, T. (2004). Three Drosophila EXT genes shape morphogen gradients through synthesis of heparan sulfate proteoglycans. Development, 131(1), 73-82. TerBush, D. R., Maurice, T., Roth, D., & Novick, P. (1996). The Exocyst is a multiprotein complex required for exocytosis in Saccharomyces cerevisiae. Tsuruhara, T., Koenig, J. H., & Ikeda, K. (1990). Synchronized endocytosis studied in the oocyte of a temperature-sensitive mutant of Drosophila melanogaster. Cell Tissue Res, 259(2), 199-207. Tu, L., Tai, W. C., Chen, L., & Banfield, D. K. (2008). Signal-mediated dynamic retention of glycosyltransferases in the Golgi. Science, 321(5887), 404-407. van Eeden, F., & St Johnston, D. (1999). The polarisation of the anterior-posterior and dorsal-ventral axes during Drosophila oogenesis. Curr Opin Genet Dev, 9(4), 396-404. Walch-Solimena, C., & Novick, P. (1999). The yeast phosphatidylinositol-4-OH kinase pik1 regulates secretion at the Golgi. Nat Cell Biol, 1(8), 523-525. Wang, J., Sun, H. Q., Macia, E., Kirchhausen, T., Watson, H., Bonifacino, J. S., & Yin, H. L. (2007). PI4P promotes the recruitment of the GGA adaptor proteins to the trans-Golgi network and regulates their recognition of the ubiquitin sorting signal. Mol Biol Cell, 18(7), 2646-2655. Wang, Y. J., Wang, J., Sun, H. Q., Martinez, M., Sun, Y. X., Macia, E., . . . Yin, H. L. (2003). Phosphatidylinositol 4 phosphate regulates targeting of clathrin adaptor AP-1 complexes to the Golgi. Cell, 114(3), 299-310. Wasserman, J. D., & Freeman, M. (1998). An autoregulatory cascade of EGF receptor signaling patterns the Drosophila egg. Cell, 95(3), 355-364. Wood, C. S., Schmitz, K. R., Bessman, N. J., Setty, T. G., Ferguson, K. M., & Burd, C. G. (2009). PtdIns4P recognition by Vps74/GOLPH3 links PtdIns 4-kinase signaling to retrograde Golgi trafficking. J Cell Biol, 187(7), 967-975. Wu, C. C., Taylor, R. S., Lane, D. R., Ladinsky, M. S., Weisz, J. A., & Howell, K. E. (2000). GMx33: a novel family of trans-Golgi proteins identified by proteomics. Traffic, 1(12), 963-975. Yeaman, C., Grindstaff, K. K., Wright, J. R., & Nelson, W. J. (2001). Sec6/8 complexes on trans-Golgi network and plasma membrane regulate late stages of exocytosis in mammalian cells. Research Support, U.S. Gov't, P.H.S.]. J Cell Biol, 155(4), 593-604. Zhang, X., Bi, E., Novick, P., Du, L., Kozminski, K. G., Lipschutz, J. H., & Guo, W. (2001). Cdc42 interacts with the exocyst and regulates polarized secretion. Research Support, U.S. Gov't, P.H.S.]. J Biol Chem, 276(50), 46745-46750. Zhang, X. M., Ellis, S., Sriratana, A., Mitchell, C. A., & Rowe, T. (2004). Sec15 is an effector for the Rab11 GTPase in mammalian cells. J Biol Chem, 279(41), 43027-43034. Zoncu, R., Perera, R. M., Balkin, D. M., Pirruccello, M., Toomre, D., & De Camilli, P. (2009). A phosphoinositide switch controls the maturation and signaling properties of APPL endosomes. Cell, 136(6), 1110-1121. Zoncu, R., Perera, R. M., Sebastian, R., Nakatsu, F., Chen, H., Balla, T., . . . De Camilli, P. V. (2007). Loss of endocytic clathrin-coated pits upon acute depletion of phosphatidylinositol 4,5-bisphosphate. Proc Natl Acad Sci U S A, 104(10), 3793-3798. | |
dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/16453 | - |
dc.description.abstract | 人類的高基氏體硫蛋白三號(Golgi phosphoprotein 3, GOLPH3),是一個主要分布於高基氏體的細胞質面的膜蛋白,其分子量為34KDa。與其他同為座落於高基氏體基質上的蛋白一樣,GOLPH3參與在物質於高機氏體基質(trans-Golgi matrix)上的順向及逆向運輸。此外也與細胞骨架交互作用以及維持高機氏體的結構。
Rotini (Y. J. Wang et al.) 為GOLPH3在果蠅中的同源物 ,先前的研究已知Rotini 協助硫醣蛋白HSPGs (Heparan Sulfate Proteoglycans)的聚合酵素 Exostosin (EXTs)蛋白在高基氏體的運輸 ,當Rotini基因在細胞中缺失時 ,會造成EXT蛋白的運輸異常,因而造成硫醣鏈(glycosaminoglycans , GAG)的多醣鏈聚合反應不完全,進而影響Hedgehog 型態決定素的表現異常以及下游訊號的缺損 。 在卵子細胞發育過程中,早期階段Rti主要表現在生殖細胞(germ cells),但到了晚期階段則主要表現在濾泡細胞內,Rti 表現位置會隨發育時期推衍而在生殖細胞以及濾泡細胞間轉換。加上先前研究得知當Rti在生殖細胞內缺失會造成胚胎扭曲及Dorsal 蛋白異常分佈表現。這些結果顯示Rti 調控胚胎背腹軸發育形成,且Rotini 可能在卵母細胞發育的過程中扮演了特定的角色。 為了更了解Rotini這個蛋白在細胞內運輸是否有所扮演其他角色,我們利用酵母菌雙雜交搜尋系統(yeast two-hybrid screen system),本論文首先證實一個參與細胞胞吐(exocytosis)相關的蛋白Sec6與Rotini蛋白有交互作用。Sec6蛋白是一個胞吐作用成員,參與將物質從高基氏體運往細胞膜的運輸。經由GST 融合蛋白沉澱實驗(GST pull down assay)確定了Rotini蛋白與Sec6蛋白有直接的交互作用。另外,在果蠅S2 細胞染色中觀察到Rotini 蛋白會與胞吐作用的成員Sec5、Sec6及Sec8部份座落在一起,進一步經由免疫沉澱實驗(Co-immunoprecipitation)證明Rotini會與胞吐作用複合體中的Sec5及Sec8有交互作用。 在果蠅卵母細胞(oocyte)中,蛋白酶Gastrulation Defective (Gd)、Snake (Snk)、Easter (Ea)及Spazle(Spz)在果蠅生殖細胞中會在靠近皮層的位置表現。當Rti在生殖細胞缺失時觀察到Snk及Snk-GFP蛋白酶無法被送至貼近皮層的位置而累積在卵母細胞中。而當Rti在生殖細胞缺失時則是觀察到Gd蛋白酶在皮層的表現量有下降的趨勢。此外,當胞吐作用成員Sec5在生殖細胞缺失時亦觀察與Rti缺失時相似的現象,會使Snk蛋白酶累積在卵母細胞中,而Gd蛋白酶則會在皮層部位表現量下降。由此推測Rti可能參與將這些蛋白酶由卵母細胞分泌至皮層的胞吐作用。 GOLPH3與4-磷脂醯肌醇 (phosphatidylinositol 4-phosphate)有直接的相互作用,且參與了細胞內物質運輸。已知4-磷脂醯肌醇主要分布於高基氏體與分泌性囊泡上。在酵母菌中,4-磷脂醯肌醇透過與Sec4p蛋白結合而參與物質由高基氏體向外運輸。因此Rti或許是以輔助者的角色來協助許多物質的運送,由Rti影響背腹軸決定分子的輸送行為,推測Rti可能藉由和4-磷脂醯肌醇的鍵結參與了許多由高基氏體向外的分泌性運輸。 | zh_TW |
dc.description.abstract | Golgi phosphoprotein 3 (GOLPH3), which is localized to the cytoplasmic face of the trans-Golgi, is a membrane protein with a molecular weight of 34 kD. GOLPH3 is one of the many proteins in the trans-Golgi matrix and is involved in anterograde and retrograde Golgi traffic, and in interactions with the cytoskeleton and maintenance of the Golgi structure.
Previous studies have proven that Rotini, a Drosophila homologue of human GOLPH3, is necessary for the retrograde trafficking of the exostosin (EXT) proteins in Golgi (Y. J. Wang et al.). EXT proteins are crucial to the glycosaminoglycans (GAG) chains biogenesis of the Heparan Sulfate Proteoglycans (HSPGs). Polymerization of GAG chains is defective when the trafficking of EXT proteins is disturbed in rti mutant cells. The morphogen Hedgehog (Hh) and its downstream signal transduction pathway are also affected in rti mutant cells. During Drosophila oogenesis, it was observed that rti is expressed in both follicle and germ cells at early stages, but mostly expressed in follicle cells at later stages when the egg chamber developed. In addition, previous data has shown that in rti loss-of-function in germline, the embryos are twisted and the distribution of Dorsal protein is affected. These results indicate that Rti participates in D/V axis formation of embryo, and it is suggested that rti could play a role in Drosophila oogenesis. To elucidate the possible roles of Rti in endo-membrane trafficking, we used yeast two-hybrid (Y2H) screen system to search for factors interacting with Rti. We found and confirmed that Sec6, an exocyst component, has physical interaction with Rti. Sec6 is involved in vesicles trafficking after post-Golgi to the plasma membrane. It was further proved that Rti has physically interacts wih Sec6 through GST pull down assay. In the staining of Drosophila S2 cell, it was shown that Rti partially co-localized with exocyst components Sec5, Sec6, and Sec8. And it was proven that Rti could be co-immunoprecipitated with Sec5 and Sec8. Through immune staining, the behaviors of serine protease cascade proteins (Gd, Snk, Ea, and Spz) in D/V development in Drosophila oogenesis was observed, and they were mostly localized in the cortex of germ cell. It was found that rti loss-of-function in germline reduced the stability of Snk and Snk-GFP proteins at the cortex of oocyte, and Snk would even clump over the space of oocyte. Additionally, It was observed that rti loss-of-function in germline reduced the localization stability of Gd proteins at the cortex of oocyte. These irregular distribution of Snk described above in rti germ-line clone is similar to that of the exocyst component Sec5E13 germ-line clone. It was observed that Sec5E13 loss-of-function in germline reduced the localization stability of Snk proteins at the cortex of oocyte and caused the Snk protein to clump over the space of oocyte. Moreover, in Sec5E13 germline clone oocyte reduced the localization stability of Gd proteins at the cortex of oocyte. From the above results, it suggests that Rti may participate in the secreted transport of these proteases to the oocyte cortex. Recently published papers have proven the direct interaction between GOLPH3, Rti homologue in human and phosphatidylinositol 4-phosphate (PtdIn4P). Furthermore, GOLPH3 were found to be involved in different trafficking pathways in the cell. PtdIn4P is specifically enriched on Golgi apparatus and secretory vesicles. In yeast PtdIn4P performs an essential role in secretory pathway through binding with Sec4p to be involved in the exocytosis of cargo from trans-Golgi network (TGN). Thereby Rti may be an auxiliary to participate in multiple trafficking pathways for different protein effectors. In other words, Rti may collaborate with PtdIn4P in the sorting of proteins into transport vesicles for exocytosis. | en |
dc.description.provenance | Made available in DSpace on 2021-06-07T18:15:37Z (GMT). No. of bitstreams: 1 ntu-101-R98b43021-1.pdf: 7743902 bytes, checksum: 95f5ef12bc9b922ace62381e55b3ee89 (MD5) Previous issue date: 2012 | en |
dc.description.tableofcontents | List of table…………………………………..…………...…..13
List of figures……………………………...…………………14 Abbreviation 17 Introduction 20 I. Oocytes……………………………………………………20 II. Dorsal-ventral axis formation in Drosophila embryo 21 1. Asymmetrically localization of Gurken activates the EGF receptor (Egfr) and determines dorsal follicle cell fates 21 2. The ventralising cascade of Drosophila 23 3. The serine protease Gd/Snk/Ea of the dorsalising pathway are needed transported to the perivitelline space 27 III. Rti, Golgi proteins evolution homologues involve Vps74p, GMx33, and GPP34 28 1. Yeast homologue of Rti is involved in protein sorting and could be a component of the retromer complex, is also required for retention of a subset of resident glycosyltransferases within Golgi 28 2. Gmx33, Homologue of Rti in mouse, is involved in several endomembrane trafficking pathways 29 3. GPP34, Homologue of Rti human, is predicted to be involved in Golgi trafficking and regulate biogenesis of mitochondria 29 IV. Rotini, the Golgi protein GOLPH3 in human, is required for HSPGs biogenesis 31 1. The rotini gene is Identificated as a segment polarity gene in Drosophila 31 2. Rotini protein does not have any conserved domain 32 3. Rotini is confirmed to be a Golgi protein 32 4. Rotini mutation exhibit the phenotypes similar to Hh signaling defects 32 5. Phenotypes in rti mutation are similar to those in HSPGs mutation 32 6. Hh expression and signaling are impaired in rotini mutation 33 7. HSPGs expression is reduced in rti mutation 33 8. Rotini mediates the HSPGs biogenesis 34 V. The exocyst complex in polarized exocytosis 35 1. The exocyst is involved in membrane addition in the cellularization of Drosophila embryo. 35 2. Exocyst is an evolutionarily conserved octameric protein complex interacting with a number of small GTPases. 36 3. Function of the exocyst in exocytosis and beyond. 38 4. Exocyst regulates polarized exocytosis by tethering post-Golgi vesicles to the bud membrane compartment, prior to their fusion. 39 5. Drosophila Sec5 is required for directing membrane traffic and the establishment of polarity within the developing oocyte 40 6. Drosophila Sec5 is present on endocytic vesicles in the oocyte 42 VI. PtdIn(4)P performs an essential role in the secretory pathway 43 1. The synthesis of phosphoinositide lipid 43 2. The spatial distributions of phosphoinositide lipids are tightly regulated 44 3. PtdIns(4)P is involved in membrane biosynthetic and vesicle budding machineries to coordinate the Golgi functions 45 4. Determinants of PtdIns(4)P levels in Golgi memebranes 45 5. Functions of PtdIns4P at the Golgi 46 6. Vesicle-mediated trafficking 46 7. Vps74P/GOLPH3--newly identified PtdIns4P effectors 47 Materials and Methods 49 1. Fly stocks and maintenance 49 2. The autosomal FLP-DFS technique 49 3. Heat shock treatment 51 4. GAL4-UAS system 51 5. Yeast two hybrid assay 52 6. GST pull-down assay 52 7. Drosophila S2 cells maintenance 54 8. Cloning constructs 57 9. Immunohistochemistry 59 10. Western blotting analysis 60 11. Membrane stripping 61 Results 63 I. The discovery of Rotini and protein interactions using the yeast two-hybrid system 64 1. The use of yeast two-hybrid assays to screen the entire genome of Drosophila 64 2. Confirmation of samples that contain in-frame AD sites 64 II. Rti interacts and colocalises with Sec6 exocyst protein 65 1. Identification of CG5341 Sec6 through yeast two-hybrid screen 65 2. Confirmation of the interactions between Rti and Sec6 in yeast K589 strain 65 3. The use of GST-pull down assay to determine the physical interaction of Rti and Sec6 66 4. The Golgi protein, Rotini, is partially colocalized with Sec6 in Drosophila S2 cells 67 III. In addition to Sec6, the other exocyst component interact with Rti 69 1. Sec5/Sec8 was also partially co-localized with Rti in S2 cell individually 69 2. Rotini co-immunoprecipitate with exocyst components, Sec5 and Sec8 70 IV. The effect of Rti involved in ventralising pathway compared with exocyst component 71 1. Rti expressed in both germline and follicle cells in early stages, but only in follicle cells after stage 9 of oogenesis. 71 2. Sec5 expressed in the membrane of both germ line and follicle cells in early stages, then concentrated on the membrane of anterolateral margin 71 3. The distribution of Grk during oogenesis 72 4. Sec5 affects the distribution of Gurken in oogenesis 73 5. Rti does not affect the distribution of Gurken in oogenesis 74 V. Rti may involved in the transportation of the serine proteases in ventralization pathway 75 1. Rti may involved in the transportation of Gd to regulate the D/V development 75 2. In the Sec5 mutant oocyte localized protease Gd was unstable 77 3. Rti may involved in the transportation of Snake to regulate the D/V development 78 4. sec5E13, sec6EX15 and rti GLC mutants have similar a Snk phenotype 80 5. Rti may be involved in Sec5 mediated exocytosis of Snk protein 80 Discussion 83 I. Rti interacted with exocyst component 83 II. Rti may involved in the transportation of the Gd and Snk in ventralising pathway 83 1. Rti affects the distributions of Gd and Snk 84 2. Sec5 affects the distributions of Gd and Snk 85 III. Immune staining of Snk and the granules accumulated in oocyte may not represent the in vivo activities of Snk protein 85 IV. Rti may transport serine proteases from oocyte to perivitelline space 86 V. The putative mechanism of Rti and PtdIn4P collaborate in secreted transport 88 VI. Future prospects 90 1. The interaction between the Rti and serine proteases 90 2. The relationships of Rti and the secreted proteins (Gd, Snk, Ea and Spz) 91 References 92 List of table Table 1.The candidates got from yeast two hybrid screen, and the function of them. ……………………………………………………………………………….101 List of figures Figure 1. The Drosophila oogenesis………………………………………………..104 Figure 2. Dorsal patterning of the eggshell is a three-stage process……………….105 Figure 3. A model for the activation of the Tollpathway……………………….......106 Figure 4. Rotini, a Golgi resided protein, which is required for HSPGs biogenesis to regulate Hh behaviors…………………………………………………………….....107 Figure 5. The Four phases of cellularization defined by Lecuit and Wieschaus…...109 Figure 6. The model for the function of the exocyst complex in tethering the secretory vesicles to the plasma membrane in yeast……………………………......110 Figure 7. Organization of the exocyst complex in the Drosophila ovaries………...112 Figure 8. Schematic illustration of the recycling route of Yolkless………………...113 Figure 9. The phosphoinositide lipid synthesis and distribution…………………..114 Figure 10. Confirmation of samples that contain in-frame AD sites………………116 Figure 11. The identification of 23-3, a fragment of Sec6, an exocyst complex components………………………………………………………………………….117 Figure 12. The confirmation of the interactions between Rti and Sec6 through yeast two-hybrid assay……………………………………………………………………118 Figure 13. Rti physically interacts with Sec6 in GST pull down assay……………119 Figure 14. Rti and Sec6 are partially colocalized in S2 cells………………………120 Figure 15. Rti and Sec5 are partially colocalized in S2 cells………………………121 Figure 16. Rti and Sec8are partially colocalized in S2 cells……………………….122 Figure 17. Immunoprecipitation staining and Western blots showing interactions between Drosophila Rti, Sec5 and Sec8 in S2 cells………………………………..123 Figure 18. The distribution of Rti during Drosophila oogenesis………………......124 Figure 19. The distribution pattern of Sec5 in during oogenesis…………………..125 Figure 20. Immunostaining and confocal microscopy of Grk in Drosophila during oogenesis……………………………………………………………………………126 Figure 21. The distribution of Grk is scattered in Sec5 loss-of-function in germline……………………………………………………………………………..127 Figure 22. The distribution of Grk is not affected when rti loss-of-function in germline……………………………………………………………………………..128 Figure 23. Gd protein distribution in the Drosophila during oogenesis……………129 Figure 24. UASp-Gd-GFP mediated by nanos-Gal4 to over-express in germline....131 Figure 25. The expression pattern of Gd in sec5E13 loss-of-function oocyte………132 Figure 26. UASp-Snk-GFP mediated by nanos-Gal4 to over-express in germline...133 Figure 27. rti loss-of-functio germline cells at stage 10 shows a reduction of localized stability of Snk-GFP proteins at the cortex of oocyte………………………………134 Figure 28. In wild type, the expression patterns of Snk in germline at the later stages of Drosophila oogenesis…………………………………………………...………..135 Figure 29. Females carrying germline mutant for sec5 produce the affected pattern of Snk antibody staining that is similar to the result from rti GLC mutation……………………………………………………………………….……136 Figure 30. Snk is scattered in early oogenesis of Sec6EX15 loss-of-function germline cell…………………………………………………………………………………..137 Figure 31. In wild type, the Snk co-localized with Sec5 in anterior corner of the oocyte……………………………………………………………………………….138 Figure 32. The distribution of Sec5 was not affected when rti loss-of-function in germline ………………………………………………………………………………………139 Figure 33. The distribution of Rti was not affected when Sec5 loss-of-function in Germline…………………………………………………………………………….140 | |
dc.language.iso | en | |
dc.title | "果蠅高基氏體蛋白GOLPH3,Rotini,與胞吐成員作用的關係探討" | zh_TW |
dc.title | The relationship between the Drosophila Golgi protein GOLPH3, Rotini, with exocyst components | en |
dc.type | Thesis | |
dc.date.schoolyear | 100-1 | |
dc.description.degree | 碩士 | |
dc.contributor.oralexamcommittee | 黃偉邦,溫進德,董桂書 | |
dc.subject.keyword | 果蠅,高機氏體蛋白,胞吐作用, | zh_TW |
dc.subject.keyword | Drosophila,Golgi protein,exocytosis, | en |
dc.relation.page | 143 | |
dc.rights.note | 未授權 | |
dc.date.accepted | 2012-02-17 | |
dc.contributor.author-college | 生命科學院 | zh_TW |
dc.contributor.author-dept | 分子與細胞生物學研究所 | zh_TW |
顯示於系所單位: | 分子與細胞生物學研究所 |
文件中的檔案:
檔案 | 大小 | 格式 | |
---|---|---|---|
ntu-101-1.pdf 目前未授權公開取用 | 7.56 MB | Adobe PDF |
系統中的文件,除了特別指名其著作權條款之外,均受到著作權保護,並且保留所有的權利。