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. 1998 Dec 1;17(23):7009–7020. doi: 10.1093/emboj/17.23.7009

dCtBP mediates transcriptional repression by Knirps, Krüppel and Snail in the Drosophila embryo.

Y Nibu 1, H Zhang 1, E Bajor 1, S Barolo 1, S Small 1, M Levine 1
PMCID: PMC1171049  PMID: 9843507

Abstract

The pre-cellular Drosophila embryo contains 10 well characterized sequence-specific transcriptional repressors, which represent a broad spectrum of DNA-binding proteins. Previous studies have shown that two of the repressors, Hairy and Dorsal, recruit a common co-repressor protein, Groucho. Here we present evidence that three different repressors, Knirps, Krüppel and Snail, recruit a different co-repressor, dCtBP. Mutant embryos containing diminished levels of maternal dCtBP products exhibit both segmentation and dorsoventral patterning defects, which can be attributed to loss of Krüppel, Knirps and Snail activity. In contrast, the Dorsal and Hairy repressors retain at least some activity in dCtBP mutant embryos. dCtBP interacts with Krüppel, Knirps and Snail through a related sequence motif, PXDLSXK/H. This motif is essential for the repression activity of these proteins in transgenic embryos. We propose that dCtBP represents a major form of transcriptional repression in development, and that the Groucho and dCtBP co-repressors mediate separate pathways of repression.

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Selected References

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

  1. Arnosti D. N., Gray S., Barolo S., Zhou J., Levine M. The gap protein knirps mediates both quenching and direct repression in the Drosophila embryo. EMBO J. 1996 Jul 15;15(14):3659–3666. [PMC free article] [PubMed] [Google Scholar]
  2. Barolo S., Levine M. hairy mediates dominant repression in the Drosophila embryo. EMBO J. 1997 May 15;16(10):2883–2891. doi: 10.1093/emboj/16.10.2883. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bier E., Jan L. Y., Jan Y. N. rhomboid, a gene required for dorsoventral axis establishment and peripheral nervous system development in Drosophila melanogaster. Genes Dev. 1990 Feb;4(2):190–203. doi: 10.1101/gad.4.2.190. [DOI] [PubMed] [Google Scholar]
  4. Brönner G., Chu-LaGraff Q., Doe C. Q., Cohen B., Weigel D., Taubert H., Jäckle H. Sp1/egr-like zinc-finger protein required for endoderm specification and germ-layer formation in Drosophila. Nature. 1994 Jun 23;369(6482):664–668. doi: 10.1038/369664a0. [DOI] [PubMed] [Google Scholar]
  5. Cai H. N., Arnosti D. N., Levine M. Long-range repression in the Drosophila embryo. Proc Natl Acad Sci U S A. 1996 Sep 3;93(18):9309–9314. doi: 10.1073/pnas.93.18.9309. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Chou T. B., Perrimon N. The autosomal FLP-DFS technique for generating germline mosaics in Drosophila melanogaster. Genetics. 1996 Dec;144(4):1673–1679. doi: 10.1093/genetics/144.4.1673. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Dubnicoff T., Valentine S. A., Chen G., Shi T., Lengyel J. A., Paroush Z., Courey A. J. Conversion of dorsal from an activator to a repressor by the global corepressor Groucho. Genes Dev. 1997 Nov 15;11(22):2952–2957. doi: 10.1101/gad.11.22.2952. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Fisher A. L., Ohsako S., Caudy M. The WRPW motif of the hairy-related basic helix-loop-helix repressor proteins acts as a 4-amino-acid transcription repression and protein-protein interaction domain. Mol Cell Biol. 1996 Jun;16(6):2670–2677. doi: 10.1128/mcb.16.6.2670. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Frasch M., Hoey T., Rushlow C., Doyle H., Levine M. Characterization and localization of the even-skipped protein of Drosophila. EMBO J. 1987 Mar;6(3):749–759. doi: 10.1002/j.1460-2075.1987.tb04817.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. González-Crespo S., Levine M. Interactions between dorsal and helix-loop-helix proteins initiate the differentiation of the embryonic mesoderm and neuroectoderm in Drosophila. Genes Dev. 1993 Sep;7(9):1703–1713. doi: 10.1101/gad.7.9.1703. [DOI] [PubMed] [Google Scholar]
  11. Gray S., Levine M. Short-range transcriptional repressors mediate both quenching and direct repression within complex loci in Drosophila. Genes Dev. 1996 Mar 15;10(6):700–710. doi: 10.1101/gad.10.6.700. [DOI] [PubMed] [Google Scholar]
  12. Gray S., Szymanski P., Levine M. Short-range repression permits multiple enhancers to function autonomously within a complex promoter. Genes Dev. 1994 Aug 1;8(15):1829–1838. doi: 10.1101/gad.8.15.1829. [DOI] [PubMed] [Google Scholar]
  13. Hanna-Rose W., Licht J. D., Hansen U. Two evolutionarily conserved repression domains in the Drosophila Kruppel protein differ in activator specificity. Mol Cell Biol. 1997 Aug;17(8):4820–4829. doi: 10.1128/mcb.17.8.4820. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Ip Y. T., Kraut R., Levine M., Rushlow C. A. The dorsal morphogen is a sequence-specific DNA-binding protein that interacts with a long-range repression element in Drosophila. Cell. 1991 Jan 25;64(2):439–446. doi: 10.1016/0092-8674(91)90651-e. [DOI] [PubMed] [Google Scholar]
  15. Ip Y. T., Park R. E., Kosman D., Bier E., Levine M. The dorsal gradient morphogen regulates stripes of rhomboid expression in the presumptive neuroectoderm of the Drosophila embryo. Genes Dev. 1992 Sep;6(9):1728–1739. doi: 10.1101/gad.6.9.1728. [DOI] [PubMed] [Google Scholar]
  16. Jiang J., Cai H., Zhou Q., Levine M. Conversion of a dorsal-dependent silencer into an enhancer: evidence for dorsal corepressors. EMBO J. 1993 Aug;12(8):3201–3209. doi: 10.1002/j.1460-2075.1993.tb05989.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Jiang J., Kosman D., Ip Y. T., Levine M. The dorsal morphogen gradient regulates the mesoderm determinant twist in early Drosophila embryos. Genes Dev. 1991 Oct;5(10):1881–1891. doi: 10.1101/gad.5.10.1881. [DOI] [PubMed] [Google Scholar]
  18. Jiménez G., Paroush Z., Ish-Horowicz D. Groucho acts as a corepressor for a subset of negative regulators, including Hairy and Engrailed. Genes Dev. 1997 Nov 15;11(22):3072–3082. doi: 10.1101/gad.11.22.3072. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Jiménez G., Pinchin S. M., Ish-Horowicz D. In vivo interactions of the Drosophila Hairy and Runt transcriptional repressors with target promoters. EMBO J. 1996 Dec 16;15(24):7088–7098. [PMC free article] [PubMed] [Google Scholar]
  20. Kasai Y., Nambu J. R., Lieberman P. M., Crews S. T. Dorsal-ventral patterning in Drosophila: DNA binding of snail protein to the single-minded gene. Proc Natl Acad Sci U S A. 1992 Apr 15;89(8):3414–3418. doi: 10.1073/pnas.89.8.3414. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Kosman D., Ip Y. T., Levine M., Arora K. Establishment of the mesoderm-neuroectoderm boundary in the Drosophila embryo. Science. 1991 Oct 4;254(5028):118–122. doi: 10.1126/science.1925551. [DOI] [PubMed] [Google Scholar]
  22. Kosman D., Small S. Concentration-dependent patterning by an ectopic expression domain of the Drosophila gap gene knirps. Development. 1997 Apr;124(7):1343–1354. doi: 10.1242/dev.124.7.1343. [DOI] [PubMed] [Google Scholar]
  23. Kraut R., Levine M. Mutually repressive interactions between the gap genes giant and Krüppel define middle body regions of the Drosophila embryo. Development. 1991 Feb;111(2):611–621. doi: 10.1242/dev.111.2.611. [DOI] [PubMed] [Google Scholar]
  24. Nambu J. R., Lewis J. O., Wharton K. A., Jr, Crews S. T. The Drosophila single-minded gene encodes a helix-loop-helix protein that acts as a master regulator of CNS midline development. Cell. 1991 Dec 20;67(6):1157–1167. doi: 10.1016/0092-8674(91)90292-7. [DOI] [PubMed] [Google Scholar]
  25. Nibu Y., Zhang H., Levine M. Interaction of short-range repressors with Drosophila CtBP in the embryo. Science. 1998 Apr 3;280(5360):101–104. doi: 10.1126/science.280.5360.101. [DOI] [PubMed] [Google Scholar]
  26. Palaparti A., Baratz A., Stifani S. The Groucho/transducin-like enhancer of split transcriptional repressors interact with the genetically defined amino-terminal silencing domain of histone H3. J Biol Chem. 1997 Oct 17;272(42):26604–26610. doi: 10.1074/jbc.272.42.26604. [DOI] [PubMed] [Google Scholar]
  27. Paroush Z., Finley R. L., Jr, Kidd T., Wainwright S. M., Ingham P. W., Brent R., Ish-Horowicz D. Groucho is required for Drosophila neurogenesis, segmentation, and sex determination and interacts directly with hairy-related bHLH proteins. Cell. 1994 Dec 2;79(5):805–815. doi: 10.1016/0092-8674(94)90070-1. [DOI] [PubMed] [Google Scholar]
  28. Paroush Z., Wainwright S. M., Ish-Horowicz D. Torso signalling regulates terminal patterning in Drosophila by antagonising Groucho-mediated repression. Development. 1997 Oct;124(19):3827–3834. doi: 10.1242/dev.124.19.3827. [DOI] [PubMed] [Google Scholar]
  29. Pazin M. J., Kadonaga J. T. What's up and down with histone deacetylation and transcription? Cell. 1997 May 2;89(3):325–328. doi: 10.1016/s0092-8674(00)80211-1. [DOI] [PubMed] [Google Scholar]
  30. Pignoni F., Baldarelli R. M., Steingrímsson E., Diaz R. J., Patapoutian A., Merriam J. R., Lengyel J. A. The Drosophila gene tailless is expressed at the embryonic termini and is a member of the steroid receptor superfamily. Cell. 1990 Jul 13;62(1):151–163. doi: 10.1016/0092-8674(90)90249-e. [DOI] [PubMed] [Google Scholar]
  31. Poortinga G., Watanabe M., Parkhurst S. M. Drosophila CtBP: a Hairy-interacting protein required for embryonic segmentation and hairy-mediated transcriptional repression. EMBO J. 1998 Apr 1;17(7):2067–2078. doi: 10.1093/emboj/17.7.2067. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Reuter R., Leptin M. Interacting functions of snail, twist and huckebein during the early development of germ layers in Drosophila. Development. 1994 May;120(5):1137–1150. doi: 10.1242/dev.120.5.1137. [DOI] [PubMed] [Google Scholar]
  33. Rivera-Pomar R., Jäckle H. From gradients to stripes in Drosophila embryogenesis: filling in the gaps. Trends Genet. 1996 Nov;12(11):478–483. doi: 10.1016/0168-9525(96)10044-5. [DOI] [PubMed] [Google Scholar]
  34. Rusch J., Levine M. Regulation of the dorsal morphogen by the Toll and torso signaling pathways: a receptor tyrosine kinase selectively masks transcriptional repression. Genes Dev. 1994 Jun 1;8(11):1247–1257. doi: 10.1101/gad.8.11.1247. [DOI] [PubMed] [Google Scholar]
  35. Sauer F., Fondell J. D., Ohkuma Y., Roeder R. G., Jäckle H. Control of transcription by Krüppel through interactions with TFIIB and TFIIE beta. Nature. 1995 May 11;375(6527):162–164. doi: 10.1038/375162a0. [DOI] [PubMed] [Google Scholar]
  36. Schaeper U., Boyd J. M., Verma S., Uhlmann E., Subramanian T., Chinnadurai G. Molecular cloning and characterization of a cellular phosphoprotein that interacts with a conserved C-terminal domain of adenovirus E1A involved in negative modulation of oncogenic transformation. Proc Natl Acad Sci U S A. 1995 Nov 7;92(23):10467–10471. doi: 10.1073/pnas.92.23.10467. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Seipel K., Georgiev O., Schaffner W. Different activation domains stimulate transcription from remote ('enhancer') and proximal ('promoter') positions. EMBO J. 1992 Dec;11(13):4961–4968. doi: 10.1002/j.1460-2075.1992.tb05603.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Small S., Arnosti D. N., Levine M. Spacing ensures autonomous expression of different stripe enhancers in the even-skipped promoter. Development. 1993 Nov;119(3):762–772. [PubMed] [Google Scholar]
  39. Small S., Blair A., Levine M. Regulation of even-skipped stripe 2 in the Drosophila embryo. EMBO J. 1992 Nov;11(11):4047–4057. doi: 10.1002/j.1460-2075.1992.tb05498.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Small S., Blair A., Levine M. Regulation of two pair-rule stripes by a single enhancer in the Drosophila embryo. Dev Biol. 1996 May 1;175(2):314–324. doi: 10.1006/dbio.1996.0117. [DOI] [PubMed] [Google Scholar]
  41. Small S., Kraut R., Hoey T., Warrior R., Levine M. Transcriptional regulation of a pair-rule stripe in Drosophila. Genes Dev. 1991 May;5(5):827–839. doi: 10.1101/gad.5.5.827. [DOI] [PubMed] [Google Scholar]
  42. Sollerbrant K., Chinnadurai G., Svensson C. The CtBP binding domain in the adenovirus E1A protein controls CR1-dependent transactivation. Nucleic Acids Res. 1996 Jul 1;24(13):2578–2584. doi: 10.1093/nar/24.13.2578. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. Stanojevic D., Small S., Levine M. Regulation of a segmentation stripe by overlapping activators and repressors in the Drosophila embryo. Science. 1991 Nov 29;254(5036):1385–1387. doi: 10.1126/science.1683715. [DOI] [PubMed] [Google Scholar]
  44. Steingrímsson E., Pignoni F., Liaw G. J., Lengyel J. A. Dual role of the Drosophila pattern gene tailless in embryonic termini. Science. 1991 Oct 18;254(5030):418–421. doi: 10.1126/science.1925599. [DOI] [PubMed] [Google Scholar]
  45. Struhl G., Johnston P., Lawrence P. A. Control of Drosophila body pattern by the hunchback morphogen gradient. Cell. 1992 Apr 17;69(2):237–249. doi: 10.1016/0092-8674(92)90405-2. [DOI] [PubMed] [Google Scholar]
  46. Sundqvist A., Sollerbrant K., Svensson C. The carboxy-terminal region of adenovirus E1A activates transcription through targeting of a C-terminal binding protein-histone deacetylase complex. FEBS Lett. 1998 Jun 12;429(2):183–188. doi: 10.1016/s0014-5793(98)00588-2. [DOI] [PubMed] [Google Scholar]
  47. Thummel C. S., Boulet A. M., Lipshitz H. D. Vectors for Drosophila P-element-mediated transformation and tissue culture transfection. Gene. 1988 Dec 30;74(2):445–456. doi: 10.1016/0378-1119(88)90177-1. [DOI] [PubMed] [Google Scholar]
  48. Turner J., Crossley M. Cloning and characterization of mCtBP2, a co-repressor that associates with basic Krüppel-like factor and other mammalian transcriptional regulators. EMBO J. 1998 Sep 1;17(17):5129–5140. doi: 10.1093/emboj/17.17.5129. [DOI] [PMC free article] [PubMed] [Google Scholar]
  49. Wharton K. A., Jr, Crews S. T. CNS midline enhancers of the Drosophila slit and Toll genes. Mech Dev. 1993 Mar;40(3):141–154. doi: 10.1016/0925-4773(93)90072-6. [DOI] [PubMed] [Google Scholar]

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