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Disease relevance of Protoplasts

  • A transferred DNA (T-DNA) tagging vector with the potential to produce dominant mutations was used with cocultured Agrobacterium tumefaciens and protoplasts to tag genes involved in the action of the plant growth substance auxin [1].
  • Efficiently regenerated single colonies from mixed multiply auxotrophic Bacillus subtilis protoplasts, fused with polyethylene glycol, reveal colonies carrying each of the parent types (biparentals) and recombinant colonies [2].
  • Coexpression of HSP70h and the p61 protein with CPm in protoplasts restricted encapsidation to the 5' approximately 630 nucleotides, which is close to the normal boundary of the bipolar virion, whereas the presence of either HSP70h or the p61 protein alone did not limit encapsidation by CPm [3].
  • One Ltk- cell in 300 to 500 was transformed to the TK+ phenotype after fusion with protoplasts carrying the chimeric plasmid pX1, which consists of pBR322 and the BamHI fragment coding for the herpes simplex virus type 1 thymidine kinase gene [4].
  • The effect of the 5'-dephosphorylated 2',5'-adenylate trimer and its 2',5'-trimer core analogs on the inhibition of tobacco mosaic virus (TMV) replication was determined in tobacco leaf discs, protoplasts, and whole tobacco plants, using infectivity tests and enzyme-linked immunosorbent assays [5].

High impact information on Protoplasts

  • Transient expression of eIF3g and L24 in plant protoplasts strongly affects TAV-mediated reinitiation activity [6].
  • A factor in conditioned medium from B. subtilis cells engineered to produce SpollR during growth triggered processing in protoplasts of B. subtilis cells that had been engineered to produce SpollGA and pro-sigma E [7].
  • Pol II-transcribed U3 RNA, containing the 5'-terminal cap different from that present in the wild-type counterpart, is packaged in transfected protoplasts into U3 snRNP precipitable with anti-fibrillarin antibodies [8].
  • A synthetic intron of arbitrary sequence but incorporating splice site consensus sequences and a high proportion of U and A nucleotides, a characteristic feature of plant introns, was efficiently spliced in protoplasts [9].
  • A complete human gamma 1 heavy chain gene (HIG1) was transferred into mouse cells by protoplast fusion [10].

Chemical compound and disease context of Protoplasts


Biological context of Protoplasts


Anatomical context of Protoplasts


Associations of Protoplasts with chemical compounds

  • Tobacco protoplasts treated with cAMP, or the adenylyl cyclase activator forskolin, no longer require auxin to divide [24].
  • Exposure to increased CO2 significantly reduced the cellular content of dimethylallyl diphosphate, the substrate for isoprene synthesis, in both leaves and leaf protoplasts [25].
  • Dark-repair of ultraviolet-induced pyrimidine dimers in the DNA of wild carrot protoplasts [26].
  • The ability of protoplasts to sustain growth in response to cytokinin glucosides persisted indefinitely after the likely disappearance of the expression vector [27].
  • A high voltage electrical pulse was applied to the protoplasts, which were then grown on filters placed over feeder layers of maize suspension cells (Black Mexican Sweet) and selected for growth in the presence of kanamycin [28].

Gene context of Protoplasts

  • Analyses of loss-of-function and ARR2-overexpressing lines as well as functional assays in protoplasts indicate an important role of ARR2 in mediating ethylene responses [29].
  • Also, a T-DNA insertion mutation at the ADL1E gene caused abnormal mitochondrial elongation that was rescued by the transient expression of ADL1C and ADL1E in protoplasts [30].
  • DREB2A domain analysis using Arabidopsis protoplasts identified a transcriptional activation domain between residues 254 and 335, and deletion of a region between residues 136 and 165 transforms DREB2A to a constitutive active form [31].
  • Using transient expression in protoplasts of BRI1 and AtSERK3 fused to cyan and yellow fluorescent green fluorescent protein variants allowed us to localize each receptor independently in vivo [32].
  • We find that ACD2 shields root protoplasts that lack chlorophyll from light- and PPIX-induced PCD [33].

Analytical, diagnostic and therapeutic context of Protoplasts

  • Here we study a chimeric integral membrane reporter protein expressed in tobacco suspension culture protoplasts whose traffic was assessed biochemically by following acquisition of complex Asn-linked glycan modifications and proteolytic processing, and whose intracellular localization was determined with confocal immunofluorescence [34].
  • By use of indirect immunofluorescence, anti-IFA stained formaldehyde-fixed onion meristematic cells and carrot protoplasts in patterns approximating those obtained with monoclonal anti-tubulins [35].
  • In immunohistochemistry and in vivo targeting experiments in Arabidopsis protoplasts, ADL1C and ADL1E appeared as numerous speckles and the two proteins colocalized [30].
  • The association of AC4 with miRNA was demonstrated by the association of A-AC4-GFP fusion protein, extracted from Arabidopsis protoplasts, with 2'-O-methyloligonucleotide complementary to miR159 (miR159*) and by the presence of miRNA with the A-AC4-GFP fusion protein after immunoprecipitation with antibody against GFP [36].
  • In this paper, we show that T-strands disappear from acetosyringone-induced A. tumefaciens within 30 min of bacterial cocultivation with tobacco protoplasts [37].


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  2. Biparental products of bacterial protoplast fusion showing unequal parental chromosome expression. Hotchkiss, R.D., Gabor, M.H. Proc. Natl. Acad. Sci. U.S.A. (1980) [Pubmed]
  3. Closterovirus bipolar virion: evidence for initiation of assembly by minor coat protein and its restriction to the genomic RNA 5' region. Satyanarayana, T., Gowda, S., Ayllón, M.A., Dawson, W.O. Proc. Natl. Acad. Sci. U.S.A. (2004) [Pubmed]
  4. High-frequency transfer of cloned herpes simplex virus type 1 sequences to mammalian cells by protoplast fusion. Sandri-Goldin, R.M., Goldin, A.L., Levine, M., Glorioso, J.C. Mol. Cell. Biol. (1981) [Pubmed]
  5. 5'-dephosphorylated 2',5'-adenylate trimer and its analogs. Inhibition of tobacco mosaic virus replication in tobacco mosaic virus-infected leaf discs, protoplasts, and intact tobacco plants. Devash, Y., Gera, A., Willis, D.H., Reichman, M., Pfleiderer, W., Charubala, R., Sela, I., Suhadolnik, R.J. J. Biol. Chem. (1984) [Pubmed]
  6. A plant viral "reinitiation" factor interacts with the host translational machinery. Park, H.S., Himmelbach, A., Browning, K.S., Hohn, T., Ryabova, L.A. Cell (2001) [Pubmed]
  7. Extracellular signal protein triggering the proteolytic activation of a developmental transcription factor in B. subtilis. Hofmeister, A.E., Londoño-Vallejo, A., Harry, E., Stragier, P., Losick, R. Cell (1995) [Pubmed]
  8. Alteration of the RNA polymerase specificity of U3 snRNA genes during evolution and in vitro. Kiss, T., Marshallsay, C., Filipowicz, W. Cell (1991) [Pubmed]
  9. The AU-rich sequences present in the introns of plant nuclear pre-mRNAs are required for splicing. Goodall, G.J., Filipowicz, W. Cell (1989) [Pubmed]
  10. Trans-acting nuclear protein responsible for induction of rearranged human immunoglobulin heavy chain gene. Maeda, H., Kitamura, D., Kudo, A., Araki, K., Watanabe, T. Cell (1986) [Pubmed]
  11. Homologous recombination in plant cells after Agrobacterium-mediated transformation. Lee, K.Y., Lund, P., Lowe, K., Dunsmuir, P. Plant Cell (1990) [Pubmed]
  12. Trypsin-induced changes in the orientation of latent ATPase in protoplast ghosts from Mycobacterium phlei. Nakagawa, H., Lee, S.H., Kalra, V.K., Brodie, A.F. J. Biol. Chem. (1977) [Pubmed]
  13. Mutanolysin-induced spheroplasts of Streptococcus mutants are true protoplasts. Siegel, J.L., Hurst, S.F., Liberman, E.S., Coleman, S.E., Bleiweis, A.S. Infect. Immun. (1981) [Pubmed]
  14. The cis-regulatory element CCACGTGG is involved in ABA and water-stress responses of the maize gene rab28. Pla, M., Vilardell, J., Guiltinan, M.J., Marcotte, W.R., Niogret, M.F., Quatrano, R.S., Pagès, M. Plant Mol. Biol. (1993) [Pubmed]
  15. Replicative transformation of the filamentous fungus Ashbya gossypii with plasmids containing Saccharomyces cerevisiae ARS elements. Wright, M.C., Philippsen, P. Gene (1991) [Pubmed]
  16. The Viviparous-1 gene and abscisic acid activate the C1 regulatory gene for anthocyanin biosynthesis during seed maturation in maize. Hattori, T., Vasil, V., Rosenkrans, L., Hannah, L.C., McCarty, D.R., Vasil, I.K. Genes Dev. (1992) [Pubmed]
  17. A feedback control element near the transcription start site of the maize Shrunken gene determines promoter activity. Maas, C., Schaal, S., Werr, W. EMBO J. (1990) [Pubmed]
  18. Auxin inducibility and developmental expression of axi 1: a gene directing auxin independent growth in tobacco protoplasts. Walden, R., Hayashi, H., Lubenow, H., Czaja, I., Schell, J. EMBO J. (1994) [Pubmed]
  19. Modulation of replication, aminoacylation and adenylation in vitro and infectivity in vivo of BMV RNAs containing deletions within the multifunctional 3' end. Bujarski, J.J., Ahlquist, P., Hall, T.C., Dreher, T.W., Kaesberg, P. EMBO J. (1986) [Pubmed]
  20. Fusion of hen erythrocytes with yeast protoplasts induced by polyethylene glycol. Ahkong, Q.F., Howell, J.I., Lucy, J.A., Safwat, F., Davey, M.R., Cocking, E.C. Nature (1975) [Pubmed]
  21. Potato sucrose transporter expression in minor veins indicates a role in phloem loading. Riesmeier, J.W., Hirner, B., Frommer, W.B. Plant Cell (1993) [Pubmed]
  22. Enzymes of glycolysis are functionally associated with the mitochondrion in Arabidopsis cells. Giegé, P., Heazlewood, J.L., Roessner-Tunali, U., Millar, A.H., Fernie, A.R., Leaver, C.J., Sweetlove, L.J. Plant Cell (2003) [Pubmed]
  23. Subcellular compartmentation of penicillin biosynthesis in Penicillium chrysogenum. The amino acid precursors are derived from the vacuole. Lendenfeld, T., Ghali, D., Wolschek, M., Kubicek-Pranz, E.M., Kubicek, C.P. J. Biol. Chem. (1993) [Pubmed]
  24. Identification and role of adenylyl cyclase in auxin signalling in higher plants. Ichikawa, T., Suzuki, Y., Czaja, I., Schommer, C., Lessnick, A., Schell, J., Walden, R. Nature (1997) [Pubmed]
  25. Increased CO2 uncouples growth from isoprene emission in an agriforest ecosystem. Rosenstiel, T.N., Potosnak, M.J., Griffin, K.L., Fall, R., Monson, R.K. Nature (2003) [Pubmed]
  26. Dark-repair of ultraviolet-induced pyrimidine dimers in the DNA of wild carrot protoplasts. Howland, G.P. Nature (1975) [Pubmed]
  27. Release of active cytokinin by a beta-glucosidase localized to the maize root meristem. Brzobohatý, B., Moore, I., Kristoffersen, P., Bako, L., Campos, N., Schell, J., Palme, K. Science (1993) [Pubmed]
  28. Genetically transformed maize plants from protoplasts. Rhodes, C.A., Pierce, D.A., Mettler, I.J., Mascarenhas, D., Detmer, J.J. Science (1988) [Pubmed]
  29. The response regulator 2 mediates ethylene signalling and hormone signal integration in Arabidopsis. Hass, C., Lohrmann, J., Albrecht, V., Sweere, U., Hummel, F., Yoo, S.D., Hwang, I., Zhu, T., Schäfer, E., Kudla, J., Harter, K. EMBO J. (2004) [Pubmed]
  30. The Arabidopsis dynamin-like proteins ADL1C and ADL1E play a critical role in mitochondrial morphogenesis. Jin, J.B., Bae, H., Kim, S.J., Jin, Y.H., Goh, C.H., Kim, D.H., Lee, Y.J., Tse, Y.C., Jiang, L., Hwang, I. Plant Cell (2003) [Pubmed]
  31. Functional Analysis of an Arabidopsis Transcription Factor, DREB2A, Involved in Drought-Responsive Gene Expression. Sakuma, Y., Maruyama, K., Osakabe, Y., Qin, F., Seki, M., Shinozaki, K., Yamaguchi-Shinozaki, K. Plant Cell (2006) [Pubmed]
  32. Heterodimerization and endocytosis of Arabidopsis brassinosteroid receptors BRI1 and AtSERK3 (BAK1). Russinova, E., Borst, J.W., Kwaaitaal, M., Caño-Delgado, A., Yin, Y., Chory, J., de Vries, S.C. Plant Cell (2004) [Pubmed]
  33. Arabidopsis ACCELERATED CELL DEATH2 modulates programmed cell death. Yao, N., Greenberg, J.T. Plant Cell (2006) [Pubmed]
  34. Integral membrane protein sorting to vacuoles in plant cells: evidence for two pathways. Jiang, L., Rogers, J.C. J. Cell Biol. (1998) [Pubmed]
  35. Monoclonal antibody to intermediate filament antigen cross-reacts with higher plant cells. Dawson, P.J., Hulme, J.S., Lloyd, C.W. J. Cell Biol. (1985) [Pubmed]
  36. MicroRNA-binding viral protein interferes with Arabidopsis development. Chellappan, P., Vanitharani, R., Fauquet, C.M. Proc. Natl. Acad. Sci. U.S.A. (2005) [Pubmed]
  37. Association of single-stranded transferred DNA from Agrobacterium tumefaciens with tobacco cells. Yusibov, V.M., Steck, T.R., Gupta, V., Gelvin, S.B. Proc. Natl. Acad. Sci. U.S.A. (1994) [Pubmed]
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