1887

Abstract

Transport of the viral genome into the nucleus is an obligatory step in the replication cycle of plant pararetro- and geminiviruses. In both these virus types, the multifunctional coat protein (CP) is thought to be involved in this process. Here, a green fluorescent protein tagging approach was used to demonstrate nuclear import of the CPs of (RTBV) and - (MYMV) in protoplasts. In both cases, at least two nuclear localization signals (NLSs) were identified and characterized. The NLSs of RTBV CP are located within both N- and C-terminal regions (residues 479KRPK/497KRK and 744KRK/758RRK), and those of MYMV CP within the N-terminal part (residues 3KR and 41KRRR). The MYMV and RTBV CP NLSs resemble classic mono- and bipartite NLSs, respectively. However, the N-terminal MYMV CP NLS and both RTBV CP NLSs show peculiarities in the number and position of basic residues. pull-down assays revealed interaction of RTBV and MYMV CPs with the nuclear import factor importin , suggesting that both CPs are imported into the nucleus via an importin -dependent pathway. The possibility that this pathway could serve for docking of virions to the nucleus is discussed.

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2005-06-01
2024-03-29
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References

  1. Adam S. A., Gerace L. 1991; Cytosolic proteins that specifically bind nuclear location signals are receptors for nuclear import. Cell 66:837–847 [CrossRef]
    [Google Scholar]
  2. Chapdelaine Y., Kirk D., Karsies A., Hohn T., Leclerc D. 2002; Mutation of capsid protein phosphorylation sites abolishes cauliflower mosaic virus infectivity. J Virol 76:11748–11752 [CrossRef]
    [Google Scholar]
  3. Cokol M., Nair R., Rost B. 2000; Finding nuclear localization signals. EMBO Rep 1:411–415 [CrossRef]
    [Google Scholar]
  4. Depienne C., Mousnier A., Leh H., Le Rouzic E., Dormont D., Benichou S., Dargemont C. 2001; Characterization of the nuclear import pathway for HIV-1 integrase. J Biol Chem 276:18102–18107 [CrossRef]
    [Google Scholar]
  5. Eckhardt S. G., Milich D. R., McLachlan A. 1991; Hepatitis B virus core antigen has two nuclear localization sequences in the arginine-rich carboxyl terminus. J Virol 65:575–582
    [Google Scholar]
  6. Gafni Y., Epel B. L. 2002; The role of host and viral proteins in intra- and inter-cellular trafficking of geminiviruses. Physiol Mol Plant Pathol 60:231–241 [CrossRef]
    [Google Scholar]
  7. Goodall G. J., Wiebauer K., Filipowicz W. 1990; Analysis of pre-mRNA processing in transfected plant protoplasts. Methods Enzymol 181:148–161
    [Google Scholar]
  8. Görlich D., Mattaj I. W. 1996; Nucleocytoplasmic transport. Science 271:1513–1518 [CrossRef]
    [Google Scholar]
  9. Görlich D., Kostka S., Kraft R., Dingwall C., Laskey R. A., Hartmann E., Prehn S. 1995a; Two different subunits of importin cooperate to recognize nuclear localization signals and bind them to the nuclear envelope. Curr Biol 5:383–392
    [Google Scholar]
  10. Görlich D., Vogel F., Mills A. D., Hartmann E., Laskey R. A. 1995b; Distinct functions for the two importin subunits in nuclear protein import. Nature 377:246–248 [CrossRef]
    [Google Scholar]
  11. Greber U. F., Fassati A. 2003; Nuclear import of viral DNA genomes. Traffic 4:136–143 [CrossRef]
    [Google Scholar]
  12. Gurevich V. V. 1996; Use of bacteriophage RNA polymerase in RNA synthesis. Methods Enzymol 275:382–397
    [Google Scholar]
  13. Haasen D., Köhler C., Neuhaus G., Merkle T. 1999; Nuclear export of proteins in plants: AtXPO1 is the export receptor for leucine-rich nuclear export signals in Arabidopsis thaliana . Plant J 20:695–705 [CrossRef]
    [Google Scholar]
  14. Harrison B. D., Swanson M. M., Fargette D. 2002; Begomovirus coat protein: serology, variation and functions. Physiol Mol Plant Pathol 60:257–271 [CrossRef]
    [Google Scholar]
  15. Hay J. M., Jones M. C., Blakebrough M. L., Dasgupta I., Davies J. W., Hull R. 1991; An analysis of the sequence of an infectious clone of rice tungro bacilliform virus, a plant pararetrovirus. Nucleic Acids Res 19:2615–2621 [CrossRef]
    [Google Scholar]
  16. Herzog E., Guerra-Peraza O., Hohn T. 2000; The rice tungro bacilliform virus gene II product interacts with the coat protein domain of the viral gene III polyprotein. J Virol 74:2073–2083 [CrossRef]
    [Google Scholar]
  17. Hull R. 1996; Molecular biology of rice tungro viruses. Annu Rev Phytopathol 34:275–297 [CrossRef]
    [Google Scholar]
  18. Imamoto N., Tachibana T., Matsubae M., Yoneda Y. 1995; A karyophilic protein forms a stable complex with cytoplasmic components prior to nuclear pore binding. J Biol Chem 270:8559–8565 [CrossRef]
    [Google Scholar]
  19. Jäkel S., Mingot J. M., Schwarzmaier P., Hartmann E., Görlich D. 2002; Importins fulfil a dual function as nuclear import receptors and cytoplasmic chaperones for exposed basic domains. EMBO J 21:377–386 [CrossRef]
    [Google Scholar]
  20. Jiang C. J., Imamoto N., Matsuki R., Yoneda Y., Yamamoto N. 1998; Functional characterization of a plant importin α homologue. Nuclear localization signal (NLS)-selective binding and mediation of nuclear import of NLS proteins in vitro . J Biol Chem 273:24083–24087 [CrossRef]
    [Google Scholar]
  21. Kalderon D., Roberts B. L., Richardson W. D., Smith A. E. 1984; A short amino acid sequence able to specify nuclear location. Cell 39:499–509 [CrossRef]
    [Google Scholar]
  22. Kann M., Sodeik B., Vlachou A., Gerlich W. H., Helenius A. 1999; Phosphorylation-dependent binding of hepatitis B virus core particles to the nuclear pore complex. J Cell Biol 145:45–55 [CrossRef]
    [Google Scholar]
  23. Karsies A., Merkle T., Szurek B., Bonas U., Hohn T., Leclerc D. 2002; Regulated nuclear targeting of cauliflower mosaic virus. J Gen Virol 83:1783–1790
    [Google Scholar]
  24. Karthikeyan A. S., Vanitharani R., Balaji V. 7 other authors 2004; Analysis of an isolate of Mungbean yellow mosaic virus (MYMV) with a highly variable DNA B component. Arch Virol 149:1643–1652
    [Google Scholar]
  25. Kotlizky G., Boulton M. I., Pitaksutheepong C., Davies J. W., Epel B. L. 2000; Intracellular and intercellular movement of maize streak geminivirus V1 and V2 proteins transiently expressed as green fluorescent protein fusions. Virology 274:32–38 [CrossRef]
    [Google Scholar]
  26. Kunik T., Palanichelvam K., Czosnek H., Citovsky V., Gafni Y. 1998; Nuclear import of the capsid protein of tomato yellow leaf curl virus (TYLCV) in plant and insect cells. Plant J 13:393–399 [CrossRef]
    [Google Scholar]
  27. 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) coat protein. J Exp Bot 50:731–732
    [Google Scholar]
  28. Leclerc D., Chapdelaine Y., Hohn T. 1999; Nuclear targeting of the cauliflower mosaic virus coat protein. J Virol 73:553–560
    [Google Scholar]
  29. Liu H., Boulton M. I., Davies J. W. 1997; Maize streak virus coat protein binds single- and double-stranded DNA in vitro . J Gen Virol 78:1265–1270
    [Google Scholar]
  30. Liu H., Boulton M. I., Thomas C. L., Prior D. A., Oparka K. J., Davies J. W. 1999; Maize streak virus coat protein is karyophyllic and facilitates nuclear transport of viral DNA. Mol Plant Microbe Interact 12:894–900 [CrossRef]
    [Google Scholar]
  31. Marmey P., Bothner B., Jacquot E., de Kochko A., Ong C. A., Yot P., Siuzdak G., Beachy R. N., Fauquet C. M. 1999; Rice tungro bacilliform virus open reading frame 3 encodes a single 37-kDa coat protein. Virology 253:319–326 [CrossRef]
    [Google Scholar]
  32. Matsuki R., Iwasaki T., Shoji K., Jiang C. J., Yamamoto N. 1998; Isolation and characterization of two importin-beta genes from rice. Plant Cell Physiol 39:879–884 [CrossRef]
    [Google Scholar]
  33. Mattaj I. W., Englmeier L. 1998; Nucleocytoplasmic transport: the soluble phase. Annu Rev Biochem 67:265–306 [CrossRef]
    [Google Scholar]
  34. Merkle T. 2001; Nuclear import and export of proteins in plants: a tool for the regulation of signalling. Planta 213:499–517 [CrossRef]
    [Google Scholar]
  35. Miller W. G., Lindow S. E. 1997; An improved GFP cloning cassette designed for prokaryotic transcriptional fusions. Gene 191:149–153 [CrossRef]
    [Google Scholar]
  36. Moroianu J., Hijikata M., Blobel G., Radu A. 1995; Mammalian karyopherin α 1 β and α 2 β heterodimers: α 1 or α 2 subunit binds nuclear localization signal and β subunit interacts with peptide repeat-containing nucleoporins. Proc Natl Acad Sci U S A 92:6532–6536 [CrossRef]
    [Google Scholar]
  37. Moroianu J., Blobel G., Radu A. 1996; Nuclear protein import: Ran-GTP dissociates the karyopherin αβ heterodimer by displacing α from an overlapping binding site on β . Proc Natl Acad Sci U S A 93:7059–7062 [CrossRef]
    [Google Scholar]
  38. Nakielny S., Dreyfuss G. 1999; Transport of proteins and RNAs in and out of the nucleus. Cell 99:677–690 [CrossRef]
    [Google Scholar]
  39. Pante N., Kann M. 2002; Nuclear pore complex is able to transport macromolecules with diameters of about 39 nm. Mol Biol Cell 13:425–434 [CrossRef]
    [Google Scholar]
  40. Pascal E., Sanderfoot A. A., Ward B. M., Medville R., Turgeon R., Lazarowitz S. G. 1994; The geminivirus BR1 movement protein binds single-stranded DNA and localizes to the cell nucleus. Plant Cell 6:995–1006 [CrossRef]
    [Google Scholar]
  41. Pooma W., Gillette W. K., Jeffrey J. L., Petty I. T. 1996; Host and viral factors determine the dispensability of coat protein for bipartite geminivirus systemic movement. Virology 218:264–268 [CrossRef]
    [Google Scholar]
  42. Qu R., Bhattacharyya M., Laco G. S. 7 other authors 1991; Characterization of the genome of rice tungro bacilliform virus: comparison with commelina yellow mottle virus and caulimoviruses. Virology 185:354–364 [CrossRef]
    [Google Scholar]
  43. Rabe B., Vlachou A., Pante N., Helenius A., Kann M. 2003; Nuclear import of hepatitis B virus capsids and release of the viral genome. Proc Natl Acad Sci U S A 100:9849–9854 [CrossRef]
    [Google Scholar]
  44. Rexach M., Blobel G. 1995; Protein import into nuclei: association and dissociation reactions involving transport substrate, transport factors, and nucleoporins. Cell 83:683–692 [CrossRef]
    [Google Scholar]
  45. Rhee Y., Gurel F., Gafni Y., Dingwall C., Citovsky V. 2000; A genetic system for detection of protein nuclear import and export. Nat Biotechnol 18:433–437 [CrossRef]
    [Google Scholar]
  46. Robbins J., Dilworth S. M., Laskey R. A., Dingwall C. 1991; Two interdependent basic domains in nucleoplasmin nuclear targeting sequence: identification of a class of bipartite nuclear targeting sequence. Cell 64:615–623 [CrossRef]
    [Google Scholar]
  47. Rojas M. R., Jiang H., Salati R., Xoconostle-Cázares B., Sudarshana M. R., Lucas W. J., Gilbertson R. L. 2001; Functional analysis of proteins involved in movement of the monopartite begomovirus, Tomato yellow leaf curl virus . Virology 291:110–125 [CrossRef]
    [Google Scholar]
  48. Rothnie H. M., Chapdelaine Y., Hohn T. 1994; Pararetroviruses and retroviruses: a comparative review of viral structure and gene expression strategies. Adv Virus Res 44:1–67
    [Google Scholar]
  49. Sanderfoot A. A., Ingham D. J., Lazarowitz S. G. 1996; A viral movement protein as a nuclear shuttle. The geminivirus BR1 movement protein contains domains essential for interaction with BL1 and nuclear localization. Plant Physiol 110:23–33 [CrossRef]
    [Google Scholar]
  50. Shoji K., Iwasaki T., Matsuki R., Miyao M., Yamamoto N. 1998; Cloning of a cDNA encoding an importin- α and down-regulation of the gene by light in rice leaves. Gene 212:279–286 [CrossRef]
    [Google Scholar]
  51. Smith A. E., Helenius A. 2004; How viruses enter animal cells. Science 304:237–242 [CrossRef]
    [Google Scholar]
  52. Szurek B., Marois E., Bonas U., Van den Ackerveken G. 2001; Eukaryotic features of the Xanthomonas type III effector AvrBs3: protein domains involved in transcriptional activation and the interaction with nuclear import receptors from pepper. Plant J 26:523–534 [CrossRef]
    [Google Scholar]
  53. Truant R., Cullen B. R. 1999; The arginine-rich domains present in human immunodeficiency virus type 1 Tat and Rev function as direct importin β -dependent nuclear localization signals. Mol Cell Biol 19:1210–1217
    [Google Scholar]
  54. Unseld S., Höhnle M., Ringel M., Frischmuth T. 2001; Subcellular targeting of the coat protein of African cassava mosaic geminivirus. Virology 286:373–383 [CrossRef]
    [Google Scholar]
  55. van Regenmortel M. H. V., Fauquet C. M., Bishop D. H. L. 8 other authors; 2000; Virus taxonomy: classification and nomenclature of viruses. In Seventh Report of the International Committee on Taxonomy of Viruses San Diego, CA, USA: Academic Press;
    [Google Scholar]
  56. Ward B. M., Lazarowitz S. G. 1999; Nuclear export in plants. Use of geminivirus movement proteins for a cell-based export assay. Plant Cell 11:1267–1276 [CrossRef]
    [Google Scholar]
  57. Weis K., Ryder U., Lamond A. I. 1996; The conserved amino-terminal domain of hSRP1 alpha is essential for nuclear protein import. EMBO J 15:1818–1825
    [Google Scholar]
  58. Whittaker G. R. 2003; Virus nuclear import. Adv Drug Deliv Rev 55:733–747 [CrossRef]
    [Google Scholar]
  59. Whittaker G. R., Helenius A. 1998; Nuclear import and export of viruses and virus genomes. Virology 246:1–23 [CrossRef]
    [Google Scholar]
  60. Yeh C. T., Liaw Y. F., Ou J. H. 1990; The arginine-rich domain of hepatitis B virus precore and core proteins contains a signal for nuclear transport. J Virol 64:6141–6147
    [Google Scholar]
  61. Zhang W., Olson N. H., Baker T. S., Faulkner L., Agbandje-McKenna M., Boulton M. I., Davies J. W., McKenna R. 2001; Structure of the maize streak virus geminate particle. Virology 279:471–477 [CrossRef]
    [Google Scholar]
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