Please use this identifier to cite or link to this item:
標題: P20/GFP融合蛋白對竹嵌紋病毒衛星核酸在細胞間移動的影響
Effect of P20/GFP fusion on cell to cell movement of Bamboo mosaic virus satellite RNA
作者: 劉光仰
Liu, Kuang-Yang
關鍵字: BaMV;竹嵌紋病毒
出版社: 生物科技學研究所
引用: 廖家德。2000。竹嵌紋病毒病徵表現之遺傳分析及載體構築。國立中興大學農業生物科技學研究所博士論文。 蔡佳芬。2003。利用 T-DNA 插入突變法建立 Nicotiana benthamiana 突變株以探討影響竹嵌紋病毒複製之寄主因子。國立中興大學生物科技學研究所碩士論文。 郭怡伶。2006。建構與分析影響竹嵌紋病毒累積之轉基因植物。國立中興大學生物科技學研究所碩士論文。 李雅倩。2005。發展轉殖植物表現竹嵌紋病毒及其衛星核酸並應用其於輔助病毒-衛星核酸交互作用之研究。國立中興大學農業生物科技學研究所博士論文。 Chang, B. Y., Lin, N. S., Liou, D. Y., Chen, J. P., Liou, G. G. & Hsu, Y. H. (1997). Subcellular localization of the 28 kDa protein of the triple-gene-block of bamboo mosaic potexvirus. J Gen Virol 78, 1175-9. Citovsky, V., Wong, M. L., Shaw, A. L., Prasad, B. V. & Zambryski, P. (1992). Visualization and characterization of tobacco mosaic virus movement protein binding to single-stranded nucleic acids. Plant Cell 4, 397-411. Cruz, S. S., Chapman, S., Roberts, A. G., Roberts, I. M., Prior, D. A. & Oparka, K. J. (1996). Assembly and movement of a plant virus carrying a green fluorescent protein overcoat. Proc Natl Acad Sci U S A 93, 6286-6290. Erhardt, M., Herzog, E., Lauber, E., Fritsch, C., Guilley, H., Jonard, G., Richards, K. & Bouzoubaa, S. (1999). Transgenic plants expressing the TGB1 protein of peanut clump virus complement movement of TGB1-defective peanut clump virus but not of TGB1-defective beet necrotic yellow vein virus. Plant Cell Rep 18, 614-619. Fedorkin, O., Solovyev, A., Yelina, N., Zamyatnin, A., Jr., Zinovkin, R., Makinen, K., Schiemann, J. & Yu Morozov, S. (2001). Cell-to-cell movement of potato virus X involves distinct functions of the coat protein. J Gen Virol 82, 449-458. Forster, R. L., Bevan, M. W., Harbison, S. A. & Gardner, R. C. (1988). The complete nucleotide sequence of the potexvirus white clover mosaic virus. Nucleic Acids Res 16, 291-303. Giesman-Cookmeyer, D. & Lommel, S. A. (1993). Alanine scanning mutagenesis of a plant virus movement protein identifies three functional domains. Plant Cell 5, 973-982. Gilbertson, R. L., Sudarshana, M., Jiang, H., Rojas, M. R. & Lucas, W. J. (2003). Limitations on geminivirus genome size imposed by plasmodesmata and virus-encoded movement protein: insights into DNA trafficking. Plant Cell 15, 2578-2591. Herzog, E., Hemmer, O., Hauser, S., Meyer, G., Bouzoubaa, S. & Fritsch, C. (1998). Identification of genes involved in replication and movement of peanut clump virus. Virology 248, 312-322. Hehnle, S., Wege, C. & Jeske, H. (2004). Interaction of DNA with the movement proteins of geminiviruses revisited. J Virol 78, 7698-7706. Huang, C., Y. Xie. & X. Zhou. (2009). Efficient virus-induced gene silencing in plants using a modified geminivirus DNA1 component. Plant Biotechnology Journal 7, 254-265. Hull, R. (1989). The movement of viruses in plant. Annu Rev Phytopathol 27, 213-240. Huisman, M. J., Linthorst, H. J., Bol, J. F. & Cornelissen, J. C. (1988). The complete nucleotide sequence of potato virus X and its homologies at the amino acid level with various plus-stranded RNA viruses. J Gen Virol 69, 1789-1798. Ivanov, K. I., Ivanov, P. A., Timofeeva, E. K., Dorokhov, Y. L. & Atabekov, J. G. (1994). The immobilized movement proteins of two tobamoviruses form stable ribonucleoprotein complexes with full-length viral genomic RNA. FEBS Lett 346, 217-220. Kawakami,S., Padgett,H.S., Hosokawa,D., Okada,Y., Beachy,R.N. & Watanabe,Y. (1999). Phosphorylation and/or presence of serine 37 in the movement protein of tomato mosaic tobamovirus is essential for intracellular localization and stability in vivo. J Virol 73, 6831-6840. Kiselyova, O. I., Yaminsky, I. V., Karger, E. M., Frolova, O. Y., Dorokhov, Y. L. & Atabekov, J. G. (2001). Visualization by atomic force microscopy of tobacco mosaic virus movement protein-RNA complexes formed in vitro. J Gen Virol 82, 1503-1508. Karpova,O.V., Ivanov,K.I., Rodionova,N.P., Dorokhov Yu,L. & Atabekov,J.G. (1997). Nontranslatability and dissimilar behavior in plants and protoplasts of viral RNA and movement protein complexes formed in vitro. Virology 230, 11-21. Kawakami,S., Hori,K., Hosokawa,D., Okada,Y. & Watanabe,Y. (2003). Defective tobamovirus movement protein lacking wild-type phosphorylation sites can be complemented by substitutions found in revertants. J Virol 77, 1452-1461. Leonard, D. A. & Zaitlin, M. (1982). A temperature-sensitive strain of tobacco mosaic virus defective in cell-to-cell movement generates an altered viral-coded protein. Virology 117, 416-424. Lee,J.Y., Taoka,K., Yoo,B.C., Ben-Nissan,G., Kim,D.J. & Lucas,W.J. (2005). Plasmodesmal-associated protein kinase in tobacco and Arabidopsis recognizes a subset of non-cell-autonomous proteins. Plant Cell 17, 2817-2831. Lin, N. S., Lee, Y. S., Lin, B. Y., Lee, C. W. & Hsu, Y. H. (1996). The open reading frame of bamboo mosaic potexvirus satellite RNA is not essential for its replication and can be replaced with a bacterial gene. Proc. Natl Acad Sci USA 93, 3138-3142. Lin, M. T., Kitajima, E. W., Cupertino, F. P. & Costa, C. L. (1977). Partial purification and some properties of bamboo mosaic virus. Phytopathology 67:1439-1443. Lin, N. S., Lin, F. Z., Huang, T. Y. & Hsu, Y. H. (1992). Genome properties of bamboo mosaic virus. Phytopathology 82, 731-734. Lin, N. S. & Hsu, Y. H. (1994). A satellite RNA associated with bamboo mosaic potexvirus. Virology 202, 707-714. Lin, M. K., Chang, B. Y., Liao, J. T., Lin, N. S. & Hsu, Y. H. (2004). Arg-16 and Arg-21 in the N-terminal region of the triple-gene-block protein 1 of bamboo mosaic virus are essential for virus movement. J Gen Virol 85, 251-259. Lin, M. K., Hu, C. C., Lin, N. S., Chang, B. Y. & Hsu, Y. H. (2006). Movement of potexviruses requires species-specific interactions among the cognate triple gene block proteins, as revealed by a trans-complementation assay based on the bamboo mosaic virus satellite RNA-mediated expression system. J Gen Virol 87, 1357-1367. Lin, N. S., Lin, B. Y., Lo, N. W., Hu, C. C., Chow, T. Y. & Hsu, Y. H. (1994). Nucleotide sequence of the genomic RNA of bamboo mosaic potexvirus. J Gen Virol 75, 2513-2518. Li, Y. I., Chen, Y. J., Hsu, Y. H. & Meng, M. (2001a). Characterization of the AdoMet-dependent guanylyltransferase activity that is associated with the N terminus of bamboo mosaic virus replicase. J Virol 75, 782-788.. Li, Y. I., Shih, T. W., Hsu, Y. H., Han, Y. T., Huang, Y. L. & Meng, M. (2001b). The helicase-like domain of plant potexvirus replicase participates in formation of RNA 5’cap structure by exhibiting RNA 5’-triphosphatase activity. J Virol 75,12114-12120. Marillonnet, S., Thoeringer, C., Kandzia, R., Klimyuk, V. & Gleba, Y. (2005). Systemic Agrobacterium tumefaciens-mediated transfection of viral replicons for efficient transient expression in plants. Nat Biotechnol 23, 718-723. McGeachy, K. D. & Barker, H. (2000). Potato mop-top virus RNA can move long distance in the absence of coat protein: evidence from resistant, transgenic plants. Mol Plant Microbe Interact 13, 125-128. Morozov, S. Yu., Lukasheva, L. I., Chernov, B. K., Skryabin, K. G. & Atabekov, J. G. (1987). Nucleotide sequence of the open reading frames adjacent to the coat protein cistron in potato virus X genome. FEBS Lett 213, 438-442. Morozov, S., Dolja, V. V. & Atabekov, J. G. (1989). Probable reassortment of genomic elements among elongated RNA-containing plant viruses. J Mol Evol 29, 52-62. Noueiry, A. O., Lucas, W. J. & Gilbertson, R. L. (1994). Two proteins of a plant DNA virus coordinate nuclear and plasmodesmal transport. Cell 76, 925-932. Osman, T. A., Thommes, P. & Buck, K. W. (1993). Localization of a single-stranded RNA-binding domain in the movement protein of red clover necrotic mosaic dianthovirus. J Gen Virol 74, 2453-2457. Ohno, T., Takamatsu, N., Meshi, T., Okada, Y., Nishiguchi, M. & Kiho, Y. (1983). Single amino acid substitution in 30K protein of TMV defective in virus transport function. Virology 131, 255-258. Petty, I. T. & Jackson, A. O. (1990). Mutational analysis of barley stripe mosaic virus RNA β. Virology 179, 712-718. Rojas, M. R., Noueiry, A. O., Lucas, W. J. & Gilbertson, R. L. (1998). Bean Dwarf mosaic geminivirus movement proteins recognize DNA in a form- and size-specific manner. Cell 95, 105-113. Ruiz, M. T., Voinnet, O. & Baulcombe, D. C. (1998). Initiation and maintenance of virus-induced gene silencing. Plant Cell 10, 937-946. Rupasov, V. V., Morozov, S., Kanyuka, K. V. & Zavriev, S. K. (1989). Partial nucleotide sequence of potato virus M RNA shows similarities to protexviruses in gene arrangement and the encoded amino acid sequences. J Gen Virol 70, 1861-1869. Stephan, D., Slabbe,r C., George, G., Ninov, V., Francis, K. P. & Burger, J. T. (2011). Visualization of plant viral suppressor silencing activity in intact leaf lamina by quantitative fluorescent imaging. Plant Methods 7, 25. Schmitt, C., Balmori, E., Jonard, G., Richards, K. E. & Guilley, H. (1992). In vitro mutagenesis of biologically active transcripts of beet necrotic yellow vein virus RNA 2: evidence that a domain of the 75-kDa readthrough protein is important for efficient virus assembly. Proc Natl Acad Sci U S A 89, 5715-5719. Sit, T. L. & AbouHaidar, M. G. (1993). Infectious RNA transcripts derived from cloned cDNA of papaya mosaic virus: effect of mutations to the capsid and polymerase proteins. J Gen Virol 74, 1133-1140. Solovyev, A. G., Savenkov, E. I., Agranovsky, A. A. & Morozov, S. Y. (1996). Comparisons of the genomic cis-elements and coding regions in RNA beta components of the hordeiviruses barley stripe mosaic virus, lychnis ringspot virus, and poa semilatent virus. Virology 219, 9-18. Soellick, T., Uhrig, J. F., Bucher, G. L., Kellmann, J. W. & Schreier, P. H. (2000). The movement protein NSm of tomato spotted wilt tospovirus (TSWV): RNA binding, interaction with the TSWV N protein, and identification of interacting plant proteins. Proc Natl Acad Sci U S A 97, 2373-2378. Skryabin, K. G., Kraev, A. S., Morozov, S., Rozanov, M. N., Chernov, B. K., Lukasheva, L. I. & Atabekov, J. G. (1988). The nucleotide sequence of potato virus X RNA. Nucleic Acids Res 16, 10929-10930. Tsai, M. S., Hsu, Y. H., & Lin, N. S. (1999). Bamboo mosaic virus satellite RNA (satBaMV)-encoded protein preferentially binds to satBaMV RNA. J Virol 73, 2703-2709. Vijayapalani, P., Chen, J. C. F., Liou, M. R., Chen, H. C., Hsu, Y. H. & Lin, N. S. (2011). Phosphorylation of bamboo mosaic virus satellite RNA (satBaMV)-encoded protein P20 downregulates the formation of satBaMV-P20 ribonucleoprotein complex. Nucleic Acids Res 40, 638-649. Waigmann, E., Ueki, S., Trutnyeva, K. & Citovsky, V. (2004). The ins and outs of non-destructive cell-to-cell and systemic movement of plant viruses. Crit Rev Plant Sci 23, 195-250. Waigmann,E., Chen,M.H., Bachmaier,R., Ghoshroy,S. & Citovsky,V. (2000). Regulation of plasmodesmal transport by phosphorylation of tobacco mosaic virus cell-to-cell movement protein. EMBO J 19, 4875-4884. Wong, S. M., Lee, K. C., Yu, H. H. & Leong, W. F. (1998). Phylogenetic analysis of triple gene block viruses based on the TGB 1 homolog gene indicates a convergent evolution. Virus Genes 16, 295-302. Wung, C. H., Hsu, Y. H., Liou, D. Y., Huang, W. C., Lin, N. S. & Chang, B. Y. (1999). Identification of the RNA-binding sites of the triple gene block protein 1 of bamboo mosaic potexvirus. J Gen Virol 80, 1119-1126. Yang, C. C., Liu J. S., Lin, C. P. & Lin, N. S. (1997). Nucleotide sequence and phylogenetic analysis of a bamboo mosaic potexvirus isolate from common bamboo (Banbusa vulgaris McClure). Bot Bull Acad Sin 38, 77-84.
Satellite RNAs (satRNAs) are subviral agents which depend on helper viruses and host factors for replication, encapsidation and movement. Bamboo mosaic virus (BaMV), a member of the genus Potexvirus, is the only potexvirus that naturally associates with a satRNA, designated satBaMV, which encodes a 20 kDa RNA-binding protein (P20) involved in efficient long-distance movement and replication of satBaMV. The aims of this study were to investigate the abilities of other non-cognate viruses in supporting the replication of satBaMV and to explore the effects of P20 phosphorylaion on the movement of satBaMV. Dimeric satBaMV transgenic plant lines expressing P20 fused with green fluorescent protein (GFP) either at N- or C- terminus were generated. BaMV and two non-cognate viruses, Potato virus X (PVX) and Foxtail mosaic virus (FoMV), were inoculated onto the transgenic plant to monitor their abilities in supporting satBaMV. Through the examination of GFP expressions in the inoculated transgenic plants, it was confirmed that the non-cognate helper FoMV marginally supports the replication of satBaMV, whereas PVX does not. Plasmids harboring GFP fused to the N- or C-terminus of P20 protein with mutations on the serine at amino acid position 11 (S11) to inhibit or mimic the phosphorylation state of P20 were constructed, and there cell-to-cell movement ability were examined in Chenopodium quinoa by fluorescent microscopy. The results revealed that both mutations of P20 reduced the cell-to-cell movement of satBaMV, corroborating the previous finding that phosphorylation of S11 is important for the expression and movement of satBaMV.

竹嵌紋病毒衛星核酸(satllite Bamboo mosaic virus RNA, satBaMV)必須依賴竹嵌紋病毒(Bamboo mosaic virus, BaMV)進行複製、包被及移動。BaMV是目前唯一在自然界被發現具有衛星核酸之馬鈴薯病毒屬之植物病毒,其satBaMV可轉譯出大小為20kDa的蛋白,稱作P20,可結合satBaMV並幫助其進行長距離之移動及複製。satBaMV載體之轉型植物系統,在竹嵌紋病毒接種後,能有效且大量地誘導外源基因的表現。為了進一步探討非同源性病毒是否具有誘導此系統中satBaMV載體的複製能力,本實驗運用兩種以satBaMV為載體之轉型植物品系satP20GFP及satGFPP20,分別是將綠色螢光蛋白(Green fluorescent protein, GFP)構築於satBaMV所含有的P20蛋白之C端及N端,再經由農桿菌(Agrobacterium)轉型系統得到轉型株。首先,為快速獲得具有同源性的F3子代轉型株,建立了以冷光螢光分析儀作為篩選satBaMV載體帶有GFP轉型株之方法。接著,進行BaMV及Potexvirus屬的狐尾草嵌紋病毒(Foxtail mosaic virus, FoMV)和馬鈴薯病毒X (Potato virus virus, PVX)之接種試驗,實驗結果顯示BaMV及FoMV測試病毒都可以放大GFP訊號,但以BaMV病毒的效果最佳。另外,為了運用GFP蛋白為報導基因來驗證P20蛋白第11個胺基酸Serine,經磷酸化後會影響satBaMV於細胞與細胞間的移動,首先將P20蛋白第11個胺基酸突變成Alanine及Aspartic acid的兩個質體pCS11A及pCS11D,同樣地將GFP構築在突變的P20蛋白之N端及C端,分別稱為pCGFPP20-S11A、pCGFPP20-S11D、pCP20GFP-S11A及pCP20GFP-S11D,將上述質體純化並接種於白藜,在接種第5天後利用螢光顯微鏡觀察GFP螢光蛋白分佈,發現上述4個選殖株之GFP蛋白螢光散佈於細胞之大小明顯小於將GFP蛋白分別構築於N端及C端之pCGFPP20及pCP20GFP選殖株,結果顯示P20蛋白的磷酸化確實會影響satBaMV於細胞與細胞間之移動。
其他識別: U0005-2008201214050100
Appears in Collections:生物科技學研究所

Show full item record

Google ScholarTM


Items in DSpace are protected by copyright, with all rights reserved, unless otherwise indicated.