1887

Journal of General Virology header logo

Volume 93, Issue 6

Mapping of functional region conferring nuclear localization and karyopherin α-binding activity of the C2 protein of bhendi yellow vein mosaic virus Free

Abstract

Bhendi yellow vein mosaic disease is caused by a complex consisting of a monopartite begomovirus associated with a β-satellite. The C2 protein of bhendi yellow vein mosaic virus (BYVMV) is a suppressor of post-transcriptional gene silencing and also functions as a transcriptional activator. To explore the molecular mechanisms of its nuclear trafficking and self-interaction, fusion proteins of fluorescent proteins with wild-type or mutated constructs of BYVMV C2 were expressed in tobacco protoplasts. Analyses revealed that the BYVMV C2 nuclear localization signal (NLS) was located in the N terminus of the protein, comprising aa 17–31 of C2. NLSs are recognized by a class of soluble transport receptors termed karyopherins α and β. The BYVMV C2 NLS was found to be necessary for this protein’s interaction with its nuclear import mediator, karyopherin α, ensuring its nuclear localization. Nevertheless, when deleted, C2 was found in both the cytoplasm and the nucleus, suggesting NLS-independent nuclear import of this protein. Homotypic interaction of BYVMV C2 was also found, which correlates with the nuclear localization needed for efficient activation of transcription.

  • Received:
  • Accepted:
  • Published Online:
© 2012 SGM
Loading

Article metrics loading...

/content/journal/jgv/10.1099/vir.0.038943-0
2012-06-01
2024-04-19
Loading full text...

Full text loading...

/deliver/fulltext/jgv/93/6/1367.html?itemId=/content/journal/jgv/10.1099/vir.0.038943-0&mimeType=html&fmt=ahah

References

  1. Baliji S., Sunter J., Sunter G. 2007; Transcriptional analysis of complementary sense genes in Spinach curly top virus and functional role of C2 in pathogenesis. Mol Plant Microbe Interact 20:194–206 [View Article][PubMed]
     [Google Scholar]
  2. Baliji S., Lacatus G., Sunter G. 2010; The interaction between geminivirus pathogenicity proteins and adenosine kinase leads to increased expression of primary cytokinin-responsive genes. Virology 402:238–247 [View Article][PubMed]
     [Google Scholar]
  3. Briddon R. W., Mansoor S., Bedford I. D., Pinner M. S., Saunders K., Stanley J., Zafar Y., Malik K. A., Markham P. G. 2001; Identification of DNA components required for induction of cotton leaf curl disease. Virology 285:234–243 [View Article][PubMed]
     [Google Scholar]
  4. Buchmann R. C., Asad S., Wolf J. N., Mohannath G., Bisaro D. M. 2009; Geminivirus AL2 and L2 proteins suppress transcriptional gene silencing and cause genome-wide reductions in cytosine methylation. J Virol 83:5005–5013 [View Article][PubMed]
     [Google Scholar]
  5. Chowda-Reddy R. V., Dong W., Felton C., Ryman D., Ballard K., Fondong V. N. 2009; Characterization of the cassava geminivirus transcription activation protein putative nuclear localization signal. Virus Res 145:270–278 [View Article][PubMed]
     [Google Scholar]
  6. Dong X., van Wezel R., Stanley J., Hong Y. 2003; Functional characterization of the nuclear localization signal for a suppressor of posttranscriptional gene silencing. J Virol 77:7026–7033 [View Article][PubMed]
     [Google Scholar]
  7. Draper J., Scott R., Armitage P., Walden R. 1988 Plant Genetic Transformation and Gene Expression: a Laboratory Manual Boston: Blackwell Science;
     [Google Scholar]
  8. Fromm M., Taylor L. P., Walbot V. 1985; Expression of genes transferred into monocot and dicot plant cells by electroporation. Proc Natl Acad Sci U S A 82:5824–5828 [View Article][PubMed]
     [Google Scholar]
  9. Gafni Y., Epel B. 2002; The role of host and viral proteins in intra- and inter-cellular trafficking of geminiviruses. Physiol Mol Plant Pathol 60:231–241 [View Article]
     [Google Scholar]
  10. Gopal P., Pravin Kumar P., Sinilal B., Jose J., Kasin Yadunandam A., Usha R. 2007; Differential roles of C4 and βC1 in mediating suppression of post-transcriptional gene silencing: evidence for transactivation by the C2 of Bhendi yellow vein mosaic virus, a monopartite begomovirus. Virus Res 123:9–18 [View Article][PubMed]
     [Google Scholar]
  11. Gutierrez C. 1999; Geminivirus DNA replication. Cell Mol Life Sci 56:313–329 [View Article][PubMed]
     [Google Scholar]
  12. Haley A., Zhan X., Richardson K., Head K., Morris B. 1992; Regulation of the activities of African cassava mosaic virus promoters by the AC1, AC2, and AC3 gene products. Virology 188:905–909 [View Article][PubMed]
     [Google Scholar]
  13. Hanley-Bowdoin L., Settlage S. B., Orozco B. M., Nagar S., Robertson D. 1999; Geminiviruses: models for plant DNA replication, transcription, and cell cycle regulation. Crit Rev Plant Sci 18:71–106 [View Article]
     [Google Scholar]
  14. Hormuzdi S. G., Bisaro D. M. 1995; Genetic analysis of beet curly top virus: examination of the roles of L2 and L3 genes in viral pathogenesis. Virology 206:1044–1054 [View Article][PubMed]
     [Google Scholar]
  15. Imai Y., Matsushima Y., Sugimura T., Terada M. 1991; A simple and rapid method for generating a deletion by PCR. Nucleic Acids Res 19:2785 [View Article][PubMed]
     [Google Scholar]
  16. Jeske H. 2009; Geminiviruses. Curr Top Microbiol Immunol 331:185–226 [View Article][PubMed]
     [Google Scholar]
  17. Jose J., Usha R. 2003; Bhendi yellow vein mosaic disease in India is caused by association of a DNA β satellite with a begomovirus. Virology 305:310–317 [View Article][PubMed]
     [Google Scholar]
  18. Kalderon D., Richardson W. D., Markham A. F., Smith A. E. 1984; Sequence requirements for nuclear location of simian virus 40 large-T antigen. Nature 311:33–38 [View Article][PubMed]
     [Google Scholar]
  19. Karpova T. S., Baumann C. T., He L., Wu X., Grammer A., Lipsky P., Hager G. L., McNally J. G. 2003; Fluorescence resonance energy transfer from cyan to yellow fluorescent protein detected by acceptor photobleaching using confocal microscopy and a single laser. J Microsc 209:56–70 [View Article][PubMed]
     [Google Scholar]
  20. Kenworthy A. K. 2001; Imaging protein–protein interactions using fluorescence resonance energy transfer microscopy. Methods 24:289–296 [View Article][PubMed]
     [Google Scholar]
  21. Kumar P., Usha R., Zrachya A., Levy Y., Spanov H., Gafni Y. 2006; Protein–protein interactions and nuclear trafficking of coat protein and betaC1 protein associated with Bhendi yellow vein mosaic disease. Virus Res 122:127–136 [View Article][PubMed]
     [Google Scholar]
  22. Kunik T., Mizrachy L., Citovsky V., Gafni Y. 1999; Characterization of a tomato karyopherin α that interacts with the Tomato Yellow Leaf Curl Virus (TYLCV) capsid protein. J Exp Bot 50:731–732 [View Article]
     [Google Scholar]
  23. Larkin M. A., Blackshields G., Brown N. P., Chenna R., McGettigan P. A., McWilliam H., Valentin F., Wallace I. M., Wilm A.other authors 2007; Clustal W and Clustal X version 2.0. Bioinformatics 23:2947–2948 [View Article][PubMed]
     [Google Scholar]
  24. Lazarowitz S. G., McGarry R. C., Barron Y. D., Carvalho M. F. 2004; Nuclear shuttling in plant cells. Symp Soc Exp Biol 56:157–176[PubMed]
     [Google Scholar]
  25. Lozano-Durán R., Bejarano E. R. 2011; Geminivirus C2 protein might be the key player for geminiviral co-option of SCF-mediated ubiquitination. Plant Signal Behav 6:999–1001 [View Article][PubMed]
     [Google Scholar]
  26. Lozano-Durán R., Rosas-Díaz T., Gusmaroli G., Luna A. P., Taconnat L., Deng X. W., Bejarano E. R. 2011; Geminiviruses subvert ubiquitination by altering CSN-mediated derubylation of SCF E3 ligase complexes and inhibit jasmonate signaling in Arabidopsis thaliana. Plant Cell 23:1014–1032 [View Article][PubMed]
     [Google Scholar]
  27. Mizrachy L., Dabush D., Levy Y., Aloni R., Altman A., Gafni Y. 2004; Cloning and characterization of the tomato karyopherin α1 gene promoter. Dev Growth Differ 46:515–522 [View Article][PubMed]
     [Google Scholar]
  28. Mosammaparast N., Pemberton L. F. 2004; Karyopherins: from nuclear-transport mediators to nuclear-function regulators. Trends Cell Biol 14:547–556 [View Article][PubMed]
     [Google Scholar]
  29. Nawaz-ul-Rehman M. S., Fauquet C. M. 2009; Evolution of geminiviruses and their satellites. FEBS Lett 583:1825–1832 [View Article][PubMed]
     [Google Scholar]
  30. Noris E., Jupin I., Accotto G. P., Gronenborn B. 1996; DNA-binding activity of the C2 protein of tomato yellow leaf curl geminivirus. Virology 217:607–612 [View Article][PubMed]
     [Google Scholar]
  31. Packialakshmi R. M., Usha R. 2011; A simple and efficient method for agroinfection of Vernonia cinerea with infectious clones of Vernonia yellow vein virus. Virus Genes 43:465–470 [View Article][PubMed]
     [Google Scholar]
  32. Saunders K., Bedford I. D., Briddon R. W., Markham P. G., Wong S. M., Stanley J. 2000; A unique virus complex causes Ageratum yellow vein disease. Proc Natl Acad Sci U S A 97:6890–6895 [View Article][PubMed]
     [Google Scholar]
  33. Saunders K., Norman A., Gucciardo S., Stanley J. 2004; The DNA β satellite component associated with ageratum yellow vein disease encodes an essential pathogenicity protein (βC1). Virology 324:37–47 [View Article][PubMed]
     [Google Scholar]
  34. Sorokin A. V., Kim E. R., Ovchinnikov L. P. 2007; Nucleocytoplasmic transport of proteins. Biochemistry (Mosc) 72:1439–1457 [View Article][PubMed]
     [Google Scholar]
  35. Sunter G., Bisaro D. M. 1992; Transactivation of geminivirus AR1 and BR1 gene expression by the viral AL2 gene product occurs at the level of transcription. Plant Cell 4:1321–1331 [View Article][PubMed]
     [Google Scholar]
  36. Sunter G., Bisaro D. M. 1997; Regulation of a geminivirus coat protein promoter by AL2 protein (TrAP): evidence for activation and derepression mechanisms. Virology 232:269–280 [View Article][PubMed]
     [Google Scholar]
  37. Trinks D., Rajeswaran R., Shivaprasad P. V., Akbergenov R., Oakeley E. J., Veluthambi K., Hohn T., Pooggin M. M. 2005; Suppression of RNA silencing by a geminivirus nuclear protein, AC2, correlates with transactivation of host genes. J Virol 79:2517–2527 [View Article][PubMed]
     [Google Scholar]
  38. Tzfira T., Tian G. W., Lacroix B., Vyas S., Li J., Leitner-Dagan Y., Krichevsky A., Taylor T., Vainstein A., Citovsky V. 2005; pSAT vectors: a modular series of plasmids for autofluorescent protein tagging and expression of multiple genes in plants. Plant Mol Biol 57:503–516 [View Article][PubMed]
     [Google Scholar]
  39. Voinnet O., Pinto Y. M., Baulcombe D. C. 1999; Suppression of gene silencing: a general strategy used by diverse DNA and RNA viruses of plants. Proc Natl Acad Sci U S A 96:14147–14152 [View Article][PubMed]
     [Google Scholar]
  40. Wang H., Hao L., Shung C.-Y., Sunter G., Bisaro D. M. 2003; Adenosine kinase is inactivated by geminivirus AL2 and L2 proteins. Plant Cell 15:3020–3032 [View Article][PubMed]
     [Google Scholar]
  41. Wang H., Buckley K. J., Yang X., Buchmann R. C., Bisaro D. M. 2005; Adenosine kinase inhibition and suppression of RNA silencing by geminivirus AL2 and L2 proteins. J Virol 79:7410–7418 [View Article][PubMed]
     [Google Scholar]
  42. Yaakov N., Levy Y., Belausov E., Gaba V., Lapidot M., Gafni Y. 2011; Effect of a single amino acid substitution in the NLS domain of tomato yellow leaf curl virus-Israel (TYLCV-IL) capsid protein (CP) on its activity and on the virus life cycle. Virus Res 158:8–11 [View Article][PubMed]
     [Google Scholar]
  43. Yang X., Baliji S., Buchmann R. C., Wang H., Lindbo J. A., Sunter G., Bisaro D. M. 2007; Functional modulation of the geminivirus AL2 transcription factor and silencing suppressor by self-interaction. J Virol 81:11972–11981 [View Article][PubMed]
     [Google Scholar]
  44. Zhang Z., Chen H., Huang X., Xia R., Zhao Q., Lai J., Teng K., Li Y., Liang L.other authors 2011; BSCTV C2 attenuates the degradation of SAMDC1 to suppress DNA methylation-mediated gene silencing in Arabidopsis. Plant Cell 23:273–288 [View Article][PubMed]
     [Google Scholar]
  45. Zhou X., Xie Y., Tao X., Zhang Z., Li Z., Fauquet C. M. 2003; Characterization of DNAβ associated with begomoviruses in China and evidence for co-evolution with their cognate viral DNA-A. J Gen Virol 84:237–247 [View Article][PubMed]
     [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jgv/10.1099/vir.0.038943-0
Loading
Mapping of functional region conferring nuclear localization and karyopherin α-binding activity of the C2 protein of bhendi yellow vein mosaic virus
J. Gen. Virol. 93, 1367 (2012); https://doi.org/10.1099/vir.0.038943-0
/content/journal/jgv/10.1099/vir.0.038943-0
/content/journal/jgv/10.1099/vir.0.038943-0
Loading

Data & Media loading...

Supplements

Supplementary material 1

PDF

Most cited Most Cited RSS feed

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error