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The Tobacco BY-2 Cell Line as a Model System to Understand in Planta Nuclear Coactivator Interactions

  • Riyaz A. Bhat
  • Richard D. Thompson
Part of the Biotechnology in Agriculture and Forestry book series (AGRICULTURE, volume 53)

Abstract

Manipulating the expression of a transgene in transient and stably transformed cells is a requirement for many functional analyses. Considerable advances in understanding gene regulation have come from the construction of chimeric genes, the design of methods for transfer of these constructs into target organisms and the study of their expression (Maniatis et al. 1987). The efficient transfer and expression of foreign DNA into intact eukaryotic cells is, therefore, a prerequisite for understanding gene expression and studying the underlying molecular mechanisms. This approach is, however, subject to the constraints of the cellular complexity of the organism under study (Harkins et al. 1990). In order to address this complexity, either one must have the capability to measure gene expression in a given cell-type within the organism, or a second alternative is to generate a population of uniform cell type and manipulate transgene expression in these cells. Established cell lines such as HeLa cells have played an important role in the basic understanding of the molecular and cellular biology of mammalian cells (David and Perrot-Rechenmann 2001). A large number of cell lines have also been obtained from various tissues and many species of higher plants. These include cell lines such as tobacco XD (Filner 1965); soybean (Keller et al. 1970) and the tobacco BY-2 cell line (Kato et al. 1972). Among these, the tobacco BY-2 cell line (Nicotiana tabacum L. cv. Bright Yellow) has shown unique characteristics and has been studied extensively (Nagata et al. 1992). The cell line is highly homogeneous and shows exceptionally high growth rates, doubling in 13–14 h under optimal conditions. The cell line can be easily synchronised following treatment with aphidicolin (a specific inhibitor of DNA polymerase α) and propyzamide (a microtubule-decomposing drug; Nagata and Kumagai 1999). As a result, BY-2 has emerged as a model plant cell line for plant cell cycle studies (Combettes et al. 1999). Furthermore, the BY-2 cell line can be easily transformed and stable transgenic calli and suspension cultures are readily obtained (David and Perrot-Rechenmann 2001). Plant protoplasts have been widely used for the past 30 years, and are undoubtedly still one of the most versatile analytical tools available. Because of its exceptionally high growth rates, BY-2 permits the isolation of large quantities of protoplasts for biochemical analyses. All these unique features make this cell line a powerful tool for plant molecular biologists.

Keywords

Histone Acetylation Nuclear Localisation Sequence Transient Gene Expression Saga Complex Histone Acetyltransferase Activity 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

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References

  1. An G (1995) Binary Ti plasmid vectors. Methods Mol Biol 44: 47–58PubMedGoogle Scholar
  2. Barlev NA, Candau R et al. (1995) Characterization of physical interactions of the putative transcriptional adaptor, ADA2, with acidic activation domains and TATA-binding protein. J Biol Chem 270 (33): 19337–19344PubMedCrossRefGoogle Scholar
  3. Berger SL, Pina B et al. (1992) Genetic isolation of ADA2: a potential transcriptional adaptor required for function of certain acidic activation domains. Cell 70 (2): 251–265PubMedCrossRefGoogle Scholar
  4. Bhat RA, Riehl M et al. (2003) Alteration of GCN5 levels in maize reveals dynamic responses to manipulating histone acetylation. Plant J 33 (3): 455–469PubMedCrossRefGoogle Scholar
  5. Brownell JE, Allis CD (1996) Special HATs for special occasions: linking histone acetylation to chromatin assembly and gene activation. Curr Opin Genet Dev 6 (2): 176–184PubMedCrossRefGoogle Scholar
  6. Brownell JE, Zhou J et al. (1996) Tetrahymena histone acetyltransferase A: a homolog to yeast Gcn5p linking histone acetylation to gene activation. Cell 84 (6): 843–851PubMedCrossRefGoogle Scholar
  7. Byrd C, Turner GC et al. (1998) The N-end rule pathway controls the import of peptides through degradation of a transcriptional repressor. EMBO J 17 (1): 269–277PubMedCrossRefGoogle Scholar
  8. Candau R, Berger SL (1996) Structural and functional analysis of yeast putative adaptors. Evidence for an adaptor complex in vivo. J Biol Chem 271 (9): 5237–5245PubMedCrossRefGoogle Scholar
  9. Combettes B, Reichheld JP et al. (1999) Study of phase-specific gene expression in synchronized tobacco cells. Methods Cell Sci 21 (2–3): 109–121PubMedCrossRefGoogle Scholar
  10. David KM, Perrot-Rechenmann C (2001) Characterization of a tobacco Bright Yellow 2 cell line expressing the tetracycline repressor at a high level for strict regulation of transgene expression. Plant Physiol 125 (4): 1548–1553PubMedCrossRefGoogle Scholar
  11. Dingwall C, Sharnick SV et al. (1982) A polypeptide domain that specifies migration of nucleoplasmin into the nucleus. Cell 30 (2): 449–458PubMedCrossRefGoogle Scholar
  12. Dunnwald M, Varshavsky A et al. (1999) Detection of transient in vivo interactions between substrate and transporter during protein translocation into the endoplasmic reticulum. Mol Biol Cell 10 (2): 329–344PubMedGoogle Scholar
  13. Filner P (1965) Semi-conservative replication of DNA in a higher plant cell. Exp Cell Res 39(1):33– 39Google Scholar
  14. Garcea RL, Alberts BM (1980) Comparative studies of histone acetylation in nucleosomes, nuclei, and intact cells. Evidence for special factors which modify acetylase action. J Biol Chem 255 (23): 11454–11463PubMedGoogle Scholar
  15. Grant PA, Sterner DE et al. (1998) The SAGA unfolds: convergence of transcription regulators in chromatin-modifying complexes. Trends Cell Biol 8 (5): 193–197PubMedCrossRefGoogle Scholar
  16. Hampsey M (1997) A SAGA of histone acetylation and gene expression. Trends Genet 13(11):427– 429Google Scholar
  17. Harkins KR, Jefferson RA et al. (1990) Expression of photosynthesis-related gene fusions is restricted by cell type in transgenic plants and in transfected protoplasts. Proc Natl Acad Sci USA 87 (2): 816–820PubMedCrossRefGoogle Scholar
  18. Hendzel MJ, Sun JM et al. (1994) Histone acetyltransferase is associated with the nuclear matrix. J Biol Chem 269 (36): 22894–22901PubMedGoogle Scholar
  19. Hettmann C, Soldati D (1999) Cloning and analysis of a Toxoplasma gondii histone acetyltransferase: a novel chromatin remodelling factor in Apicomplexan parasites. Nucleic Acids Res 27 (22): 4344–4352PubMedCrossRefGoogle Scholar
  20. Jach G, Binot E et al. (2001) Use of red fluorescent protein from Discosoma sp. (dsRED) as a reporter for plant gene expression. Plant J 28 (4): 483–491PubMedCrossRefGoogle Scholar
  21. Johnsson N, Varshavsky A (1994) Split ubiquitin as a sensor of protein interactions in vivo. Proc Natl Acad Sci USA 91 (22): 10340–10344PubMedCrossRefGoogle Scholar
  22. Kato K, Matsumoto T et al. (1972) Liquid suspension culture of tobacco cells. In: Terui G (ed) Fermentation technology today. Society of fermentation technology, Osaka, pp 689–695Google Scholar
  23. Keller WA, Harvey B et al. (1970) Plant protoplasts for use in somatic cell hybridization. Nature 226 (242): 280–282PubMedCrossRefGoogle Scholar
  24. Kuo MH, Zhou J et al. (1998) Histone acetyltransferase activity of yeast Gcn5p is required for the activation of target genes in vivo. Genes Dev 12 (5): 627–639PubMedCrossRefGoogle Scholar
  25. Lane T, Ibanez C et al. (1990) Transformation by v-myb correlates with trans-activation of gene expression. Mol Cell Biol 10 (6): 2591–2598PubMedGoogle Scholar
  26. Lehming N (2001) Mechanisms of transcriptional repression. Max Planck Institute for Plant Breeding Research, Cologne, pp 155–157Google Scholar
  27. Loidl P (1988) Towards an understanding of the biological function of histone acetylation. FEBS Lett 227 (2): 91–95PubMedCrossRefGoogle Scholar
  28. Lopez-Rodas G, Perez-Ortin JE et al. (1985) Partial purification and properties of two histone acetyltransferases from the yeast, Saccharomyces cerevisiae. Arch Biochem Biophys 239 (1): 184–190PubMedCrossRefGoogle Scholar
  29. Maniatis T, Goodbourn S et al. (1987) Regulation of inducible and tissue-specific gene expression. Science 236 (4806): 1237–1245PubMedCrossRefGoogle Scholar
  30. Marcus GA, Silverman N et al. (1994) Functional similarity and physical association between GCN5 and ADA2: putative transcriptional adaptors. EMBO J 13 (20): 4807–4815PubMedGoogle Scholar
  31. Nagata T, Kumagai F (1999) Plant cell biology through the window of the highly synchronized tobacco BY-2 cell line. Methods Cell Sci 21 (2–3): 123–127PubMedCrossRefGoogle Scholar
  32. Nagata T, Okada K et al. (1981) Delivery of tobacco mosaic virus RNA into plant protoplasts by reverse-phase evaporation vesicles (liposomes). Mol Gen Genet 184: 161–165Google Scholar
  33. Nagata T, Nemoto A, Hasezawa S (1992) Tobacco BY-2 cell line as the “HeLâ cell in the cell biology of higher plants. Int Rev Cytol 132: 1–30CrossRefGoogle Scholar
  34. Negrutiu I, Shillito R et al. (1987) Hybrid genes in the analysis of transformation conditions. Plant Mol Biol 8: 363–373CrossRefGoogle Scholar
  35. Oliva R, Bazett-Jones DP et al. (1990) Histone hyperacetylation can induce unfolding of the nucleosome core particle. Nucleic Acids Res 18 (9): 2739–2747PubMedCrossRefGoogle Scholar
  36. Ornaghi P, Ballario P et al. (1999) The bromodomain of Gcn5p interacts in vitro with specific residues in the N terminus of histone H4. J Mol Biol 287 (1): 1–7CrossRefGoogle Scholar
  37. Peters R, Lang I et al. (1986) Fluorescence microphotolysis to measure nucleocytoplasmic transport in vivo and in vitro. Biochem Soc Trans 14 (5): 821–822PubMedGoogle Scholar
  38. Pollard KJ, Peterson CL (1997) Role for ADA/GCN5 products in antagonizing chromatin-mediated transcriptional repression. Mol Cell Biol 17 (11): 6212–6222PubMedGoogle Scholar
  39. Potrykus I (1990) Gene transfer methods for plants and cell cultures. Ciba Found Symp 154:198– 208; discussion 208–212Google Scholar
  40. Rojo-Niersbach E, Morley D et al. (2000) A new method for the selection of protein interactions in mammalian cells. Biochem J 348 (3): 585–590PubMedCrossRefGoogle Scholar
  41. Sheen J, Hwang S et al. (1995) Green-fluorescent protein as a new vital marker in plant cells. Plant J 8 (5): 777–784PubMedCrossRefGoogle Scholar
  42. Silver PA (1991) How proteins enter the nucleus. Cell 64 (3): 489–497PubMedCrossRefGoogle Scholar
  43. Silverman N, Agapite J et al. (1994) Yeast ADA2 protein binds to the VP16 protein activation domain and activates transcription. Proc Natl Acad Sci USA 91 (24): 11665–11668PubMedCrossRefGoogle Scholar
  44. Sterner DE, Wang X et al. (2002) The SANT domain of Ada2 is required for normal acetylation of histones by the yeast SAGA complex. J Biol Chem 277 (10): 8178–8186PubMedCrossRefGoogle Scholar
  45. Stockinger EJ, Mao Y et al. (2001) Transcriptional adaptor and histone acetyltransferase proteins in Arabidopsis and their interactions with CBF1, a transcriptional activator involved in cold-regulated gene expression. Nucleic Acids Res 29 (7): 1524–1533PubMedCrossRefGoogle Scholar
  46. Töpfer R, Schell J et al. (1988) Versatile cloning vectors for transient gene expression and direct gene transfer in plant cells. Nucleic Acids Res 16 (17): 8725PubMedCrossRefGoogle Scholar
  47. Varshavsky A (1996) The N-end rule: functions, mysteries, uses. Proc Natl Acad Sci USA 93 (22): 12142–12149PubMedCrossRefGoogle Scholar
  48. Varshavsky A (1997) The ubiquitin system. Trends Biochem Sci 22 (10): 383–387PubMedCrossRefGoogle Scholar
  49. Wang L, Liu L et al. (1998) Critical residues for histone acetylation by Gcn5, functioning in Ada and SAGA complexes, are also required for transcriptional function in vivo. Genes Dev 12 (5): 640–653PubMedCrossRefGoogle Scholar
  50. Wiegand RC, Brutlag DL (1981) Histone acetylase from Drosophila melanogaster specific for H4. J Biol Chem 256 (9): 4578–4583PubMedGoogle Scholar
  51. Wittke S, Lewke N et al. (1999) Probing the molecular environment of membrane proteins in vivo. Mol Biol Cell 10 (8): 2519–2530PubMedGoogle Scholar
  52. Zhao LJ, Padmanabhan R (1988) Nuclear transport of adenovirus DNA polymerase is facilitated by interaction with preterminal protein. Cell 55 (6): 1005–1015PubMedCrossRefGoogle Scholar

Copyright information

© Springer-Verlag Berlin Heidelberg 2004

Authors and Affiliations

  • Riyaz A. Bhat
    • 1
  • Richard D. Thompson
    • 2
  1. 1.Max-Planck-Institut für ZüchtungsforschungKölnGermany
  2. 2.INRA-URGELP Legume UnitDijon CédexFrance

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