Skip to main content
PMC home page
Nucleic Acids Research logoLink to Nucleic Acids Research
. 1997 Aug 15;25(16):3345–3353. doi: 10.1093/nar/25.16.3345

Multiple isoforms of GAGA factor, a critical component of chromatin structure.

C Benyajati 1, L Mueller 1, N Xu 1, M Pappano 1, J Gao 1, M Mosammaparast 1, D Conklin 1, H Granok 1, C Craig 1, S Elgin 1
PMCID: PMC146888  PMID: 9241251

Abstract

The GAGA transcription factor of Drosophila melanogaster is ubiquitous and plays multiple roles. Characterization of cDNA clones and detection by domain- specific antibodies has revealed that the 70-90 kDa major GAGA species are encoded by two open reading frames producing GAGA factor proteins of 519 amino acids (GAGA-519) and 581 amino acids (GAGA-581), which share a common N-terminal region that is linked to two different glutamine-rich C-termini. Purified recombinant GAGA-519 and GAGA-581 proteins can form homomeric complexes that bind specifically to a single GAGA sequence in vitro. The two GAGA isoforms also function similarly in transient transactivation assays in tissue culture cells and in chromatin remodeling experiments in vitro . Only GAGA-519 protein accumulates during the first 6 h of embryogenesis. Thereafter, both GAGA proteins are present in nearly equal amounts throughout development; in larval salivary gland nuclei they colocalize completely to specific regions along the euchromatic arms of the polytene chromosomes. Coimmunoprecipitation of GAGA-519 and GAGA-581 from crude nuclear extracts and from mixtures of purified recombinant proteins, indicates direct interactions. We suggest that homomeric complexes of GAGA-519 may function during early embryogenesis; both homomeric and heteromeric complexes of GAGA-519 and GAGA-581 may function later.

Full Text

The Full Text of this article is available as a PDF (504.5 KB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Ayer S., Benyajati C. Conserved enhancer and silencer elements responsible for differential Adh transcription in Drosophila cell lines. Mol Cell Biol. 1990 Jul;10(7):3512–3523. doi: 10.1128/mcb.10.7.3512. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Ayer S., Benyajati C. The binding site of a steroid hormone receptor-like protein within the Drosophila Adh adult enhancer is required for high levels of tissue-specific alcohol dehydrogenase expression. Mol Cell Biol. 1992 Feb;12(2):661–673. doi: 10.1128/mcb.12.2.661. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Ayer S., Walker N., Mosammaparast M., Nelson J. P., Shilo B. Z., Benyajati C. Activation and repression of Drosophila alcohol dehydrogenase distal transcription by two steroid hormone receptor superfamily members binding to a common response element. Nucleic Acids Res. 1993 Apr 11;21(7):1619–1627. doi: 10.1093/nar/21.7.1619. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Bardwell V. J., Treisman R. The POZ domain: a conserved protein-protein interaction motif. Genes Dev. 1994 Jul 15;8(14):1664–1677. doi: 10.1101/gad.8.14.1664. [DOI] [PubMed] [Google Scholar]
  5. Becker P. B. Drosophila chromatin and transcription. Semin Cell Biol. 1995 Aug;6(4):185–190. doi: 10.1006/scel.1995.0026. [DOI] [PubMed] [Google Scholar]
  6. Becker P. B., Tsukiyama T., Wu C. Chromatin assembly extracts from Drosophila embryos. Methods Cell Biol. 1994;44:207–223. doi: 10.1016/s0091-679x(08)60915-2. [DOI] [PubMed] [Google Scholar]
  7. Benyajati C., Ewel A., McKeon J., Chovav M., Juan E. Characterization and purification of Adh distal promoter factor 2, Adf-2, a cell-specific and promoter-specific repressor in Drosophila. Nucleic Acids Res. 1992 Sep 11;20(17):4481–4489. doi: 10.1093/nar/20.17.4481. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Bhat K. M., Farkas G., Karch F., Gyurkovics H., Gausz J., Schedl P. The GAGA factor is required in the early Drosophila embryo not only for transcriptional regulation but also for nuclear division. Development. 1996 Apr;122(4):1113–1124. doi: 10.1242/dev.122.4.1113. [DOI] [PubMed] [Google Scholar]
  9. Biggin M. D., Tjian R. Transcription factors that activate the Ultrabithorax promoter in developmentally staged extracts. Cell. 1988 Jun 3;53(5):699–711. doi: 10.1016/0092-8674(88)90088-8. [DOI] [PubMed] [Google Scholar]
  10. Bond-Matthews B., Davidson N. Transcription from each of the Drosophila act5C leader exons is driven by a separate functional promoter. Gene. 1988;62(2):289–300. doi: 10.1016/0378-1119(88)90566-5. [DOI] [PubMed] [Google Scholar]
  11. Brown N. H., Kafatos F. C. Functional cDNA libraries from Drosophila embryos. J Mol Biol. 1988 Sep 20;203(2):425–437. doi: 10.1016/0022-2836(88)90010-1. [DOI] [PubMed] [Google Scholar]
  12. Bunch T. A., Grinblat Y., Goldstein L. S. Characterization and use of the Drosophila metallothionein promoter in cultured Drosophila melanogaster cells. Nucleic Acids Res. 1988 Feb 11;16(3):1043–1061. doi: 10.1093/nar/16.3.1043. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Chen W., Zollman S., Couderc J. L., Laski F. A. The BTB domain of bric à brac mediates dimerization in vitro. Mol Cell Biol. 1995 Jun;15(6):3424–3429. doi: 10.1128/mcb.15.6.3424. [DOI] [PMC free article] [PubMed] [Google Scholar] [Retracted]
  14. Clark R. F., Wagner C. R., Craig C. A., Elgin S. C. Distribution of chromosomal proteins in polytene chromosomes of Drosophila. Methods Cell Biol. 1991;35:203–227. doi: 10.1016/s0091-679x(08)60574-9. [DOI] [PubMed] [Google Scholar]
  15. Croston G. E., Kerrigan L. A., Lira L. M., Marshak D. R., Kadonaga J. T. Sequence-specific antirepression of histone H1-mediated inhibition of basal RNA polymerase II transcription. Science. 1991 Feb 8;251(4994):643–649. doi: 10.1126/science.1899487. [DOI] [PubMed] [Google Scholar]
  16. Dong S., Zhu J., Reid A., Strutt P., Guidez F., Zhong H. J., Wang Z. Y., Licht J., Waxman S., Chomienne C. Amino-terminal protein-protein interaction motif (POZ-domain) is responsible for activities of the promyelocytic leukemia zinc finger-retinoic acid receptor-alpha fusion protein. Proc Natl Acad Sci U S A. 1996 Apr 16;93(8):3624–3629. doi: 10.1073/pnas.93.8.3624. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Driever W., Nüsslein-Volhard C. The bicoid protein is a positive regulator of hunchback transcription in the early Drosophila embryo. Nature. 1989 Jan 12;337(6203):138–143. doi: 10.1038/337138a0. [DOI] [PubMed] [Google Scholar]
  18. Emili A., Greenblatt J., Ingles C. J. Species-specific interaction of the glutamine-rich activation domains of Sp1 with the TATA box-binding protein. Mol Cell Biol. 1994 Mar;14(3):1582–1593. doi: 10.1128/mcb.14.3.1582. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Farkas G., Gausz J., Galloni M., Reuter G., Gyurkovics H., Karch F. The Trithorax-like gene encodes the Drosophila GAGA factor. Nature. 1994 Oct 27;371(6500):806–808. doi: 10.1038/371806a0. [DOI] [PubMed] [Google Scholar]
  20. Gill G., Pascal E., Tseng Z. H., Tjian R. A glutamine-rich hydrophobic patch in transcription factor Sp1 contacts the dTAFII110 component of the Drosophila TFIID complex and mediates transcriptional activation. Proc Natl Acad Sci U S A. 1994 Jan 4;91(1):192–196. doi: 10.1073/pnas.91.1.192. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Gilmour D. S., Thomas G. H., Elgin S. C. Drosophila nuclear proteins bind to regions of alternating C and T residues in gene promoters. Science. 1989 Sep 29;245(4925):1487–1490. doi: 10.1126/science.2781290. [DOI] [PubMed] [Google Scholar]
  22. Granok H., Leibovitch B. A., Shaffer C. D., Elgin S. C. Chromatin. Ga-ga over GAGA factor. Curr Biol. 1995 Mar 1;5(3):238–241. doi: 10.1016/s0960-9822(95)00048-0. [DOI] [PubMed] [Google Scholar]
  23. Hakes D. J., Dixon J. E. New vectors for high level expression of recombinant proteins in bacteria. Anal Biochem. 1992 May 1;202(2):293–298. doi: 10.1016/0003-2697(92)90108-j. [DOI] [PubMed] [Google Scholar]
  24. Jackson S. P., Tjian R. O-glycosylation of eukaryotic transcription factors: implications for mechanisms of transcriptional regulation. Cell. 1988 Oct 7;55(1):125–133. doi: 10.1016/0092-8674(88)90015-3. [DOI] [PubMed] [Google Scholar]
  25. Kerrigan L. A., Croston G. E., Lira L. M., Kadonaga J. T. Sequence-specific transcriptional antirepression of the Drosophila Krüppel gene by the GAGA factor. J Biol Chem. 1991 Jan 5;266(1):574–582. [PubMed] [Google Scholar]
  26. Lu Q., Wallrath L. L., Granok H., Elgin S. C. (CT)n (GA)n repeats and heat shock elements have distinct roles in chromatin structure and transcriptional activation of the Drosophila hsp26 gene. Mol Cell Biol. 1993 May;13(5):2802–2814. doi: 10.1128/mcb.13.5.2802. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. O'Brien T., Wilkins R. C., Giardina C., Lis J. T. Distribution of GAGA protein on Drosophila genes in vivo. Genes Dev. 1995 May 1;9(9):1098–1110. doi: 10.1101/gad.9.9.1098. [DOI] [PubMed] [Google Scholar]
  28. Pascal E., Tjian R. Different activation domains of Sp1 govern formation of multimers and mediate transcriptional synergism. Genes Dev. 1991 Sep;5(9):1646–1656. doi: 10.1101/gad.5.9.1646. [DOI] [PubMed] [Google Scholar]
  29. Pedone P. V., Ghirlando R., Clore G. M., Gronenborn A. M., Felsenfeld G., Omichinski J. G. The single Cys2-His2 zinc finger domain of the GAGA protein flanked by basic residues is sufficient for high-affinity specific DNA binding. Proc Natl Acad Sci U S A. 1996 Apr 2;93(7):2822–2826. doi: 10.1073/pnas.93.7.2822. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Raff J. W., Kellum R., Alberts B. The Drosophila GAGA transcription factor is associated with specific regions of heterochromatin throughout the cell cycle. EMBO J. 1994 Dec 15;13(24):5977–5983. doi: 10.1002/j.1460-2075.1994.tb06943.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Shopland L. S., Hirayoshi K., Fernandes M., Lis J. T. HSF access to heat shock elements in vivo depends critically on promoter architecture defined by GAGA factor, TFIID, and RNA polymerase II binding sites. Genes Dev. 1995 Nov 15;9(22):2756–2769. doi: 10.1101/gad.9.22.2756. [DOI] [PubMed] [Google Scholar]
  32. Soeller W. C., Oh C. E., Kornberg T. B. Isolation of cDNAs encoding the Drosophila GAGA transcription factor. Mol Cell Biol. 1993 Dec;13(12):7961–7970. doi: 10.1128/mcb.13.12.7961. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Soeller W. C., Poole S. J., Kornberg T. In vitro transcription of the Drosophila engrailed gene. Genes Dev. 1988 Jan;2(1):68–81. doi: 10.1101/gad.2.1.68. [DOI] [PubMed] [Google Scholar]
  34. Stott K., Blackburn J. M., Butler P. J., Perutz M. Incorporation of glutamine repeats makes protein oligomerize: implications for neurodegenerative diseases. Proc Natl Acad Sci U S A. 1995 Jul 3;92(14):6509–6513. doi: 10.1073/pnas.92.14.6509. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Tsukiyama T., Becker P. B., Wu C. ATP-dependent nucleosome disruption at a heat-shock promoter mediated by binding of GAGA transcription factor. Nature. 1994 Feb 10;367(6463):525–532. doi: 10.1038/367525a0. [DOI] [PubMed] [Google Scholar]
  36. Tsukiyama T., Wu C. Purification and properties of an ATP-dependent nucleosome remodeling factor. Cell. 1995 Dec 15;83(6):1011–1020. doi: 10.1016/0092-8674(95)90216-3. [DOI] [PubMed] [Google Scholar]
  37. Vinson C. R., LaMarco K. L., Johnson P. F., Landschulz W. H., McKnight S. L. In situ detection of sequence-specific DNA binding activity specified by a recombinant bacteriophage. Genes Dev. 1988 Jul;2(7):801–806. doi: 10.1101/gad.2.7.801. [DOI] [PubMed] [Google Scholar]
  38. Wall G., Varga-Weisz P. D., Sandaltzopoulos R., Becker P. B. Chromatin remodeling by GAGA factor and heat shock factor at the hypersensitive Drosophila hsp26 promoter in vitro. EMBO J. 1995 Apr 18;14(8):1727–1736. doi: 10.1002/j.1460-2075.1995.tb07162.x. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Nucleic Acids Research are provided here courtesy of Oxford University Press

RESOURCES