ORIGINAL ARTICLE
Analysis of the interaction between Tomato torrado virus proteins using the yeast two-hybrid system
 
More details
Hide details
1
Interdepartmental Laboratory of Molecular Biology, Institute of Plant Protection – National Research Institute, Władysława Węgorka 20, 60-318 Poznań, Poland
CORRESPONDING AUTHOR
Aleksandra Obrępalska-Stęplowska
Interdepartmental Laboratory of Molecular Biology, Institute of Plant Protection – National Research Institute, Władysława Węgorka 20, 60-318 Poznań, Poland
Submission date: 2013-10-22
Acceptance date: 2013-10-30
 
Journal of Plant Protection Research 2013;53(4):416–423
KEYWORDS
TOPICS
ABSTRACT
Ten years ago for the first time the new picorna-like virus species – Tomato torrado virus (ToTV) – was found and described on tomato plants. The isolates of this pathogen were reported in Europe, America, and Oceania including Australia. Because of its unique biological and molecular features, ToTV was classified to the new genus Torradovirus, in the Secoviridae family. In Poland, three isolates: Wal’03, Kra, and Ros ToTV were identified on greenhouse tomato cultivars. At present, the biology and the genome structure of this virus are characterised. But there is no data extending beyond the bioinformatics analyses about the function of viral proteins, polyproteins, and non-coding sequences, as well as possible interactions between viral, host and vector factors that may be important for the infection process, encapsidation, transport in plants, and transmission. In this study, we have undertaken a search for the possible protein-protein interaction of ToTV encoded proteins using the yeast two-hybrid (Y2H) system. The viral genome fragments covering full sequences for nine known proteins of ToTV were amplified using specific primers with characteristic recombination sites. This process enabled the construction of basic entry clones for each protein that further facilitated manipulations with prepared constructs using Gateway technology. Two-hybrid assays were performed in the yeast strain and tested interactions of ToTV proteins were analysed in several combinations using auxotrophy markers. Our analyses did not reveal the presence of interactions between ToTV domains. Surprisingly, no interactions were found in the case of various CP subunits as well as between CP subunits and 3A protein, that in some virus families are known to play a role in viral life cycle. This role includes virion assembly or cell-to-cell transport. The lack of interactions may be a result of the limitation of this experimental system, or suggest that these proteins may interact indirectly, or require the presence of genomic RNAs or some host factors.
CONFLICT OF INTEREST
The authors have declared that no conflict of interests exist.
 
REFERENCES (34)
1.
Alfaro-Fernández A., Córdoba-Sellés C., Cebrián M.C., Sánchez-Navarro J.A., Espino A., Martín R., Jordá C. 2007. First report of Tomato torrado virus in Tomato in the Canary Islands, Spain. Plant Dis. 91 (8): 1060.
 
2.
Alfaro-Fernández A., Córdoba-Sellés C., Cebrián M.C., Herrera-Vásquez J.A. 2008. First report of Tomato torrado virus on weed hosts in Spain. Plant Dis. 92 (5): 831.
 
3.
Alfaro-Fernández A., Bese G., Córdoba-Sellés C., Cebrián M.C., Herrera-Vásquez J.A., Forray A., Jordá C. 2009. First report of Tomato torrado virus infecting Tomato in Hungary. Plant Dis. 93 (5): 554.
 
4.
Alfaro-Fernández A., Del Carmen M., Córdoba-Sellés C., Miguel Juárez J.A., Herrera-Vásquez J.A., Sánchez-Navarro J.A., Del Carmen Cebrián M., Font M.I., Jordá C. 2010. Occurrence and geographical distribution of the ‘Torrado’ disease in Spain. J. Phytopathol. 158 (7–8): 457–469.
 
5.
Amari K., Gonzalez-Ibeas P., Gómez R.N., Sempere M.A., Sanchez-Pina M.A., Aranda M.A., Moriones E., Blanca J., Hernandez- Gallardo M. D., Anastasio G. 2008. Tomato torrado virus is Transmitted by Bemisia tabaci and infects pepper and eggplant in addition to tomato. Plant Dis. 92 (7): 1139.
 
6.
Auerbach D., Stagljar I., 2005. Yeast Two-Hybrid Protein–Protein Interaction Networks. p.19–31. In: “Proteomics and Protein-Protein Interactions. Biology, Chemistry, Bioinformatics, and Drug Design”. Protein Reviews 3 (G. Waksman, ed.). Springer Science Business Media, Inc., Singapore, 323 pp
 
7.
Bol J.F. 1999. Alfalfa mosaic virus and ilarviruses: involvement of coat protein in multiple steps of the replication cycle. J. Gen. Virol. 80: 1089–1102.
 
8.
Budziszewska M., Obrepalska-Steplowska A., Wieczorek P., Pospieszny H. 2008. The nucleotide sequence of a Polish isolate of Tomato torrado virus. Virus Genes 37 (3): 400–406.
 
9.
Budziszewska M., Wieczorek P., Obrępalska-Stęplowska A. 2011. The study of the differences in the accumulation of Kra and Wal’03 isolates of Tomato torrado virus in host plant. [Badanie różnic w akumulacji izolatów Kra i Wal’03 wirusa nekrozy pomidora w roślinie gospodarza]. Prog. Plant Prot./Post. Ochr. Roślin 51 (4): 1612–1616.
 
10.
Causier B., Davies B. 2002. Analysing protein-protein interactions with the yeast two-hybrid system. Plant Mol. Biol. 50 (6): 855–870.
 
11.
Davino S., Bivona L., Iacono G., Davino M. 2010. First report of Tomato torrado virus infecting tomato in Italy. Plant Dis. 94 (9): 1172.
 
12.
De Graaff M., Man in’t Veld M.R., Jaspars E.M. 1995. In vitro evidence that the coat protein of alfalfa mosaic virus plays a direct role in the regulation of plus and minus RNA synthesis: implications for the life cycle of alfalfa mosaic virus. Virology 20 (208): 583–589.
 
13.
EPPO 2009. EPPO Allert list. Internet resource http://www.eppo.org/ [Accessed: August 17, 2013].
 
14.
Gambley C.F., Thomas J,E., Persley D.M., Hall B.H. 2010. First report of Tomato torrado virus on tomato from Australia. Plant Dis. 94 (4): 486.
 
15.
Herranz M.C., Pallas V., Aparicio F. 2012. Multifunctional roles of the N-terminal basic motif of Alfalfa mosaic virus coat protein: nucleolar/cytoplasmic shuttling, modulation of RNA-binding activity and virion formation. Mol. Plant Microbe Interact. 25 (8): 1093–1103.
 
16.
Herrera-Vasquez J.A., Alfaro-Fernández A., Cordoba-Selles M.C., Cebrian M.C., Font M.I., Jorda C. 2009. First report of Tomato torrado virus infecting tomato in single and mixed infections with Cucumber mosaic virus in Panama. Plant Dis. 93 (2): 198.
 
17.
Hull R. 2001. Matthews’ Plant Virology. 4th ed. Academic Press, San Diego, 1001 pp.
 
18.
Mateu M.G. 2013. Assembly, stability and dynamics of virus capsids. Arch. Biochem. Biophys. 531 (1–2): 65–79.
 
19.
Pospieszny H., Borodynko N., Hasiów B., Obrępalska-Stęplowska A., Budziszewska M. 2008. Tomato torrado virus – new pathogen of tomato. [Wirus nekrozy pomidora (Tomato torrado virus) – nowy patogen pomidora]. Prog. Plant Prot./ Post. Ochr. Roślin 48 (3): 1086–1096.
 
20.
Pospieszny H., Borodynko N., Hasiów B., Obrępalska-Stęplowska A., Budziszewska M. 2007b. Tomato torrado virus – New virus transmitted by greenhouse whitefly (Trialeurodes vaporariorum) in Poland. p. 69. In: Proc. 10th Int. Plant Virus Epidemiology Symposium, Icrisat, India, 15–19 October 2007, 178 pp.
 
21.
Pospieszny H., Borodynko N., Obrepalska-Steplowska A., Hasiow B. 2007a. The first report of Tomato torrado virus in Poland. Plant Dis. 91 (10): 1364.
 
22.
Pospieszny H., Budziszewska M., Hasiów-Jaroszewska B., Obrępalska-Stęplowska A., Borodynko N. 2010. Biological and molecular characterization of Polish isolates of Tomato torrado virus. J. Phytopathol. 158 (1): 56–62.
 
23.
Pospieszny H., Hasiów-Jaroszewska B., Rymelska N., Borodynko N. 2012. Using the IC real-time RT-PCR technique for the detection of Tomato torrado virus (ToTV) in tomato seedling from infected seeds. [Zastosowanie techniki IC real-time RT-PCR do wykrywania wirusa nekrozy pomidora (ToTV) w siewkach z porażonych nasion]. Prog. Plant Prot./Post. Ochr. Roślin 52 (3): 515–517.
 
24.
Rajagopala S.V., Hughes K.T., Uetz P. 2009 Benchmarking yeast two-hybrid systems using the interactions of bacterial motility proteins. Proteomics 9 (23): 5296–5302.
 
25.
Ritzenthaler Ch. 2011. Parallels and distinctions in the direct cell-to-cell spread of the plant and animal viruses. Curr Opin Virol. 1 (5): 403–409.
 
26.
Sambrook J., Fritsch E.F., Maniatis T. 2001. Molecular Cloning. A Laboratory Manual, 3rd ed. Cold Spring Harbor Laboratory Press. New York, USA.
 
27.
Sanfacon H., Wellink J., Le Gall O., Karasev A., Van der Vlugt R. 2009. Secoviridae: a proposed family of plant viruses within the order Picornavirales that combines the families Sequiviridae and Comoviridae, the unassigned genera Cheravirus and Sadwavirus, and the proposed genus Torradovirus. Arch. Virol. 154 (5): 899–907.
 
28.
Schoelz J.E., Harries P.A., Nelson R.S. 2011. Intracellular transport of plant viruses: finding the door out of the cell. Mol. Plant 4 (5): 813–831.
 
29.
Tate J., Liljas L., Scotti P., Christian P., Lin T., Johnson J.E. 1999. The crystal structure of cricket paralysis virus: the first view of a new virus family. Nat. Struct. Biol. 6: 765–774.
 
30.
Van der Heijden M.W., Carette J.E., Reinhoud P.J., Haegi A., Bol J.F. 2001. Alfalfa mosaic virus replicase proteins P1 and P2 interact and colocalize at the vacuolar membrane. J. Virol. 75 (4): 1879–1887.
 
31.
Verbeek M., Dullemans A.M., van den Heuvel J.F.J., Maris P.C., van der Flugt R.A.A. 2007. Identification and characterization of Tomato torrado virus, a new picornalike virus from tomato. Arch Virol. 152: 881–990.
 
32.
Verbeek M., Dullemans A.M. 2012. First Report of Tomato torrado virus Infecting Tomato in Colombia. Plant Dis. 96 (4): 592.
 
33.
Verdin E., Gognalons P., Wipf-Scheibel C., Bornard I., Ridray G., Schoen L., Lecoq H. 2009. First Report of Tomato torrado virus in Tomato Crops in France. Plant Dis. 93 (12): 1352.
 
34.
Wieczorek P., Obrępalska-Stęplowska A. 2013. Multiplex RT-PCR reaction for simultaneous detection of Tomato torrado virus and Pepino mosaic virus co-infecting Solanum lycopersicum. J. Plant Prot. Res. 53 (3): 289–294.
 
eISSN:1899-007X
ISSN:1427-4345