Virology Journal | |
Cocksfoot mottle virus coat protein is dispensable for the systemic infection | |
Erkki Truve2  Jelena Gerassimenko2  Cecilia Sarmiento2  Kristjan Kamsol2  Allan Olspert1  | |
[1] Current address: Department of Plant Sciences, University of Cambridge, Downing Street, Cambridge CB2 3EA, UK;Department of Gene Technology, Tallinn University of Technology, Akadeemia tee 15, 12618 Tallinn, Estonia | |
关键词: Coat protein; RNA silencing suppressor; Virus movement; Sobemovirus; | |
Others : 812267 DOI : 10.1186/1743-422X-11-19 |
|
received in 2013-11-17, accepted in 2014-01-27, 发布年份 2014 | |
【 摘 要 】
Background
The Sobemovirus genome consists of polycistronic single-stranded positive-sense RNA. The first ORF encodes P1, a suppressor of RNA silencing required for virus movement. The coat protein (CP) is expressed from the 3′ proximal ORF3 via subgenomic RNA. In addition to its structural role, the CP of some sobemoviruses has been reported to be required for systemic movement and to interact with P1. The aim of this study was to analyse the role of Cocksfoot mottle virus (CfMV) CP in the suppression of RNA silencing and virus movement.
Methods
Agrobacterium-mediated transient expression method was used for testing CfMV CP capacity to suppress RNA silencing. CP substitution and deletion mutants were generated to examine the role of this protein in CfMV infection, using three host plants (oat, barley and wheat). The viral movement was characterised with CfMV expressing EGFP fused to the C-terminus of CP.
Results
In the current study we show that CfMV CP is an additional RNA silencing suppressor. Interestingly, we observed that all CP mutant viruses were able to infect the three tested host plants systemically, although usually with reduced accumulation. CfMV expressing EGFP was detected in epidermal and mesophyll cells of inoculated leaves. Although EGFP fluorescence was not detected in upper leaves, some plants displayed CfMV symptoms. Analysis of the upper leaves revealed that the viruses had lost the EGFP sequence and sometimes also most of the CP gene.
Conclusions
The present study demonstrates that CfMV CP suppresses RNA silencing but, surprisingly, is dispensable for systemic movement. Thus, CfMV does not move as virion in the tested host plants. The composition of the movement RNP complex remains to be elucidated.
【 授权许可】
2014 Olspert et al.; licensee BioMed Central Ltd.
【 预 览 】
Files | Size | Format | View |
---|---|---|---|
20140709082010828.pdf | 1833KB | download | |
Figure 8. | 63KB | Image | download |
Figure 7. | 178KB | Image | download |
Figure 5. | 48KB | Image | download |
Figure 4. | 80KB | Image | download |
Figure 3. | 67KB | Image | download |
Figure 2. | 73KB | Image | download |
Figure 1. | 174KB | Image | download |
【 图 表 】
Figure 1.
Figure 2.
Figure 3.
Figure 4.
Figure 5.
Figure 7.
Figure 8.
【 参考文献 】
- [1]Truve E, Fargette D: Genus Sobemovirus. In Virus Taxonomy Classification and Nomenclature of Viruses: ninth Report of the International Committee on Taxonomy of Viruses. Edited by King AMQ, Adams MJ, Carstens EB, Lefkowitz EJ. London: Elsevier; 2012:1185-1189.
- [2]Mäkinen K, Næss V, Tamm T, Truve E, Aaspõllu A, Saarma M: The putative replicase of the cocksfoot mottle sobemovirus is translated as a part of the polyprotein by -1 ribosomal frameshift. Virology 1995, 207:566-571.
- [3]Abad-Zapatero C, Abdel-Meguid SS, Johnson JE, Leslie AG, Rayment I, Rossmann MG, Suck D, Tsukihara T: Structure of southern bean mosaic virus at 2.8 Å resolution. Nature 1980, 286:33-39.
- [4]Hermodson MA, Abad-Zapatero C, Abdel-Meguid SS, Pundak S, Rossmann MG, Tremaine JH: Amino acid sequence of southern bean mosaic virus coat protein and its relation to the three-dimensional structure of the virus. Virology 1982, 119:133-149.
- [5]Plevka P, Tars K, Zeltins A, Balke I, Truve E, Liljas L: The three-dimensional structure of ryegrass mottle virus at 2.9 Å resolution. Virology 2007, 369:364-374.
- [6]Qu C, Liljas L, Opalka N, Brugidou C, Yeager M, Beachy RN, Fauquet CM, Johnson JE, Lin T: 3D domain swapping modulates the stability of members of an icosahedral virus group. Structure 2000, 8:1095-1103.
- [7]Rossmann MG, Abad-Zapatero C, Hermodson MA, Erickson JW: Subunit interactions in southern bean mosaic virus. J Mol Biol 1983, 166:37-73.
- [8]Tars K, Zeltins A, Liljas L: The three-dimensional structure of cocksfoot mottle virus at 2.7 Å resolution. Virology 2003, 310:287-297.
- [9]Lee SK, Hacker DL: In vitro analysis of an RNA binding site within the N-terminal 30 amino acids of the southern cowpea mosaic virus coat protein. Virology 2001, 286:317-327.
- [10]Satheshkumar PS, Lokesh GL, Murthy MR, Savithri HS: The role of arginine-rich motif and beta-annulus in the assembly and stability of Sesbania mosaic virus capsids. J Mol Biol 2005, 353:447-458.
- [11]Tamm T, Truve E: RNA-binding activities of cocksfoot mottle sobemovirus proteins. Virus Res 2000, 66:197-207.
- [12]Lee SK, Dabney-Smith C, Hacker DL, Bruce BD: Membrane activity of the southern cowpea mosaic virus coat protein: the role of basic amino acids, helix-forming potential, and lipid composition. Virology 2001, 291:299-310.
- [13]Olspert A, Paves H, Toomela R, Tamm T, Truve E: Cocksfoot mottle sobemovirus coat protein contains two nuclear localization signals. Virus Genes 2010, 40:423-431.
- [14]Brugidou C, Holt C, Yassi MN, Zhang S, Beachy R, Fauquet C: Synthesis of an infectious full-length cDNA clone of rice yellow mottle virus and mutagenesis of the coat protein. Virology 1995, 206:108-115.
- [15]Sivakumaran K, Fowler BC, Hacker DL: Identification of viral genes required for cell-to-cell movement of southern bean mosaic virus. Virology 1998, 252:376-386.
- [16]Fuentes AL, Hamilton RI: Failure of long-distance movement of southern bean mosaic virus in a resistant host is correlated with lack of normal virion formation. J Gen Virol 1993, 74:1903-1910.
- [17]Opalka N, Brugidou C, Bonneau C, Nicole M, Beachy RN, Yeager M, Fauquet C: Movement of rice yellow mottle virus between xylem cells through pit membranes. Proc Natl Acad Sci USA 1998, 95:3323-3328.
- [18]Chowdhury SR, Savithri HS: Interaction of Sesbania mosaic virus movement protein with the coat protein – implications for viral spread. FEBS J 2011, 278:257-272.
- [19]Meier M, Paves H, Olspert A, Tamm T, Truve E: P1 protein of Cocksfoot mottle virus is indispensable for the systemic spread of the virus. Virus Genes 2006, 32:321-326.
- [20]Sarmiento C, Gomez E, Meier M, Kavanagh TA, Truve E: Cocksfoot mottle virus P1 suppresses RNA silencing in Nicotiana benthamiana and Nicotiana tabacum. Virus Res 2007, 123:95-99.
- [21]Lu R, Folimonov A, Shintaku M, Li WX, Falk BW, Dawson WO, Ding SW: Three distinct suppressors of RNA silencing encoded by a 20-kb viral RNA genome. Proc Natl Acad Sci USA 2004, 101:15742-15747.
- [22]Senshu H, Yamaji Y, Minato N, Shiraishi T, Maejima K, Hashimoto M, Miura C, Neriya Y, Namba S: A dual strategy for the suppression of host antiviral silencing: two distinct suppressors for viral replication and viral movement encoded by potato virus M. J Virol 2011, 85:10269-10278.
- [23]Kouassi NK, Chen L, Sire C, Bangratz-Reyser M, Beachy RN, Fauquet CM, Brugidou C: Expression of rice yellow mottle virus coat protein enhances virus infection in transgenic plants. Arch Virol 2006, 151:2111-2122.
- [24]Callaway AS, George CG, Lommel SA: A Sobemovirus coat protein gene complements long-distance movement of a coat protein-null Dianthovirus. Virology 2004, 330:186-195.
- [25]Mäkinen K, Tamm T, Næss V, Truve E, Puurand Ü, Munthe T, Saarma M: Characterization of cocksfoot mottle sobemovirus genomic RNA and sequence comparison with related viruses. J Gen Virol 1995, 76:2817-2825.
- [26]Logemann J, Schell J, Willmitzer L: Improved method for the isolation of RNA from plant tissues. Anal Biochem 1987, 163:16-20.
- [27]Tamm T, Mäkinen K, Truve E: Identification of genes encoding for the cocksfoot mottle virus proteins. Arch Virol 1999, 144:1557-1567.
- [28]Bendahmane A, Farnham G, Moffett P, Baulcombe DC: Constitutive gain-of-function mutants in a nucleotide binding site-leucine rich repeat protein encoded at the Rx locus of potato. Plant J 2002, 32:195-204.
- [29]Hamilton CM, Frary A, Lewis C, Tanksley SD: Stable transfer of intact high molecular weight DNA into plant chromosomes. Proc Natl Acad Sci USA 1996, 93:9975-9979.
- [30]Hamilton A, Voinnet O, Chappell L, Baulcombe D: Two classes of short interfering RNA in RNA silencing. EMBO J 2002, 21:4671-4679.
- [31]Akbergenov R, Si-Ammour A, Blevins T, Amin I, Kutter C, Vanderschuren H, Zhang P, Gruissem W, Meins F Jr, Hohn T, Pooggin MM: Molecular characterization of geminivirus-derived small RNAs in different plant species. Nucleic Acids Res 2006, 34:462-471.
- [32]Szittya G, Molnar A, Silhavy D, Hornyik C, Burgyan J: Short defective interfering RNAs of tombusviruses are not targeted but trigger post-transcriptional gene silencing against their helper virus. Plant Cell 2002, 14:359-372.