Abstract
Transfections into Schneider’s Drosophila line 2 (abbreviated SL2 or S2) derived from Drosophila embryos (1) have been used to analyze activation properties of mammalian transcription factors (Table 1), to identify activation and inhibitory domains, as well as to investigate specific protein-protein interactions in vivo. SL2 cells are particularly suited to this task because they are devoid of many ubiquitous mammalian transcription factor activities and thus their transcriptional properties can be investigated in the absence of interference by endogenous factors. The usage of SL2 cells, as a host for studying the structure and function of heterologous, mammalian transcription factors, was originally reported by the group of R. Tjian to identify functional domains of AP-2 (2), Sp1 (3,4) and CTF/NF-I (5).
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Schneider, I. (1972) Cell lines derived from late embryonic stages of Drosophila melanogaster. J. Embryol. Exp. Morphol. 27, 353–365.
Williams, T., Admon, A., Luscher, B., and Tjian, R. (1988) Cloning and expression of AP-2, a cell-type-specific transcription factor that activates inducible enhancer elements. Genes Dev. 2, 1557–1569.
Courey, A. J. and Tjian, R. (1988) Analysis of Sp1 in vivo reveals multiple transcriptional domains, including a novel glutamine-rich activation motif. Cell 55, 887–898.
Courey, A. J., Holtzman, D. A., Jackson, S. P., and Tjian, R. (1989) Synergistic activation by the glutamine-rich domains of human transcription factor Sp1. Cell 59, 827–836.
Mermod, N., O’Neill, E. A., Kelly, T. J., and Tjian, R. (1989) The proline-rich transcriptional activator of CTF/NF-I is distinct from the replication and DNA binding domain. Cell 58, 741–753.
DiNocera, P. P. and Dawid, I. B. (1983) Transient expression of genes introduced into cultured cells of Drosophila. Proc. Natl. Acad. Sci. USA 80, 7095–7098.
Ausubel, F. M., Brent, R., Kingston, R. E., Moore, D. D., Seidman, J. G., Smith, J. A., and Struhl, K., eds. (1997) Current Protocols in Molecular Biology, Wiley, New York.
Andrews, N. C. and Faller, D. V. (1991) A rapid micropreparation technique for extraction of DNA-binding proteins from limiting numbers of mammalian cells. Nucleic Acids Res. 19, 2499.
Pascal, E. and Tjian, R. (1991) Different activation domains of Sp1 govern formation of multimers and mediate transcriptional synergism. Genes Dev. 5, 1646–1656.
Yoshinaga, S. K. and Yamamoto, K. R. (1991) Signaling and regulation by a mammalian glucocorticoid receptor in Drosophila cells. Mol. Endocrinol. 5, 844–853.
Gregory, R. C., Taxman, D. J., Seshasayee, D., Kensinger, M. H., Bieker, J. J., and Wojchowski, D. M. (1996) Functional interaction of GATA1 with erythroid Krüppel-like factor and Sp1 at defined erythroid promoters. Blood 87, 1793–1801.
Merika, M. and Orkin, S. H. (1995) Functional synergy and physical interactions of the erythroid transcription factor GATA-1 with the Krüppel family proteins Sp1 and EKLF. Mol. Cell. Biol. 15, 2437–2447.
Galera, P., Musso, M., Ducy, P., and Karsenty, G. (1994) c-Krox, a transcriptional regulator of type I collagen gene expression, is preferentially expressed in skin. Proc. Natl. Acad. Sci. USA 91, 9372–9376.
Chavrier, P., Vesque, C., Galliot, B., Vigneron, M., Dollé, P., Duboule, D., and Charnay, P. (1990) The segment-specific gene Krox-20 encodes a transcription factor with binding sites in the promoter region of the Hox-1,4 gene. EMBO J. 9, 1209–1218.
Hagen, G., Müller, S., Beato, M., and Suske, G. (1994) Sp1-mediated transcriptional activation is repressed by Sp3. EMBO J. 13, 3843–3851.
Hagen, G., Dennig, J., Preiβ, A., Beato, M., and Suske, G. (1995) Functional analyses of the transcription factor Sp4 reveal properties distinct from Sp1 and Sp3. J. Biol. Chem. 270, 24,989–24,994.
Kobayashi, A., Sogawa, K., and Fujii, K. Y. (1996) Cooperative interaction between AhR.Arnt and Sp1 for the drug-inducible expression of CYP1A1 gene. J. Biol. Chem. 271, 12,310–12,316.
Seto, E., Lewis, B., and Shenk, T. (1993) Interaction between transcription factors Sp1 and YY1. Nature 365, 462–464.
Xu, J., Thompson, K. L., Shephard, L. B., Hudson, L. G., and Gill, G. N. (1993) T3 receptor suppression of Sp1-dependent transcription from the epidermal growth factor receptor promoter via overlapping DNA-binding sites. J. Biol. Chem. 268, 16,065–16,073.
Nehls, M. C., Grapilon, M. L., and Brenner, D. A. (1992) NF-I/Sp1 switch elements regulate collagen alpha 1(I) gene expression. DNA Cell. Biol. 11, 443–452.
Fischer, K.-D., Haese, A., and Nowock, J. (1993) Cooperation of GATA-1 and Sp1 can result in synergistic transcriptional activation or interference. J. Biol. Chem. 268, 23,915–23,923.
Gegonne, A., Bosselut, R., Bailly, R. A., and Ghysdael, J. (1993) Synergistic activation of the HTLV1 LTR ets-responsive region by transcription factors ets1 and Sp1. EMBO J. 12, 1169–1178.
Chen, X., Farmer, G., Zhu, H., Prywes, R., and Prives, C. (1993) Cooperative DNA binding of p53 with TFIID (TBP): a possible mechanism for transcriptional activation. Genes Dev. 7, 1837–1849.
Kerr, L. D., Ransone, L. J., Wamsley, P., Schmitt, M. J., Boyer, T. G., Zhou, Q., Berk, A. J., and Verma, I. M. (1993) Association between proto-oncoprotein Rel and TATA-binding protein mediates transcriptional activation by NF-kappa B. Nature 365, 412–419.
Yieh, L., Sanchez, H. B., and Osborne, T. F. (1995) Domains of transcription factor Sp1 required for synergistic activation with sterol regulatory element binding protein 1 of low density lipoprotein receptor promoter. Proc. Natl. Acad. Sci. USA 92, 6102–6106.
Roulet, E., Armentero, M. T., Krey, G., Corthesy, B., Dreyer, C., Mermod, N., and Wahli, W. (1995) Regulation of the DNA-binding and transcriptional activities of Xenopus laevis NFI-X by a novel C-terminal domain. Mol. Cell. Biol. 15, 5552–5562.
Lin, S.-Y., Black, A. R., Kostic, D., Pajovic, S., Hoover, C. N., and Azizkhan, J. C. (1996) Cell cycle-regulated association of E2F and Sp1 is related to their functional interaction. Mol. Cell. Biol. 16, 1668–1675.
Lee, Y. H., Williams, S. C., Baer, M., Sterneck, E., Gonzalez, F. J., and Johnson, P. F. (1997) The ability of C/EBPβ but not C/EBPα to synergize with an Sp1 protein is specified by the leucine zipper and activation domain. Mol. Cell. Biol. 17, 2038–2047.
Farmer, G., Friedlander, P., Colgan, J., Manley, J. L., and Prives, C. (1996) Transcriptional repression by p53 involves molecular interactions distinct from those with the TATA box binding protein. Nucl. Acids Res. 24, 4281–4288.
Trivedi, A., Vilalta, A., Gopalan, S., and Johnson, D. L. (1996) TATA-binding protein is limiting for both TATA-containing and TATA-lacking RNA polymerase III promoters in Drosophila cells. Mol. Cell. Biol. 16, 6909–6916.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2000 Humana Press Inc., Totowa, NJ
About this protocol
Cite this protocol
Suske, G. (2000). Transient Transfection of Schneider Cells in the Study of Transcription Factors. In: Tymms, M.J. (eds) Transcription Factor Protocols. Methods in Molecular Biology™, vol 130. Humana Press. https://doi.org/10.1385/1-59259-686-X:175
Download citation
DOI: https://doi.org/10.1385/1-59259-686-X:175
Publisher Name: Humana Press
Print ISBN: 978-0-89603-573-7
Online ISBN: 978-1-59259-686-7
eBook Packages: Springer Protocols