Abstract
During development of the Drosophila adult appendage precursors, the larval imaginal disks, the decapentaplegic (dpp) gene is expressed in a stripe just anterior to the anterior/posterior (A/P) compartment boundary. Here, we investigate the genetic controls that lead to production of this stripe. We extend previous observations on leaky engrailed (en) mutations by showing that mutant clones completely lacking both en and invected (inv) activity ectopically express dpp-lacZ reporter genes in the posterior compartment, where dpp activity ordinarily is repressed. Similarly, patched (ptc) is also ectopically expressed in such posterior compartment en(-)inv(-) null clones. In contrast, these en(-)inv(-) clones exhibit loss of hedgehog (hh) expression. We suggest that the absence of dpp expression in the posterior compartment is due to direct repression by en. Ubiquitious expression of en in imaginal disks, produced by a hs-en construct, eliminates the expression of dpp-lacZ in its normal A/P boundary stripe. We identify three in vitro Engrailed binding sites in one of our dpp-lacZ reporter gene. Mutagenesis of these Engrailed binding sites results in ectopic expression of this reporter gene, but does not alter the normal stripe of expression at the A/P boundary. We propose that the en-hh-ptc regulatory loop that is responsible for segmental expression of wingless in the embryo is reutilized in imaginal disks to create a stripe of dpp expression along the A/P compartment boundary.
Full Text
The Full Text of this article is available as a PDF (11.1 MB).
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Blackman R. K., Sanicola M., Raftery L. A., Gillevet T., Gelbart W. M. An extensive 3' cis-regulatory region directs the imaginal disk expression of decapentaplegic, a member of the TGF-beta family in Drosophila. Development. 1991 Mar;111(3):657–666. doi: 10.1242/dev.111.3.657. [DOI] [PubMed] [Google Scholar]
- Campbell G., Weaver T., Tomlinson A. Axis specification in the developing Drosophila appendage: the role of wingless, decapentaplegic, and the homeobox gene aristaless. Cell. 1993 Sep 24;74(6):1113–1123. doi: 10.1016/0092-8674(93)90732-6. [DOI] [PubMed] [Google Scholar]
- Capdevila J., Estrada M. P., Sánchez-Herrero E., Guerrero I. The Drosophila segment polarity gene patched interacts with decapentaplegic in wing development. EMBO J. 1994 Jan 1;13(1):71–82. doi: 10.1002/j.1460-2075.1994.tb06236.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Coleman K. G., Poole S. J., Weir M. P., Soeller W. C., Kornberg T. The invected gene of Drosophila: sequence analysis and expression studies reveal a close kinship to the engrailed gene. Genes Dev. 1987 Mar;1(1):19–28. doi: 10.1101/gad.1.1.19. [DOI] [PubMed] [Google Scholar]
- Crick F. H., Lawrence P. A. Compartments and polyclones in insect development. Science. 1975 Aug 1;189(4200):340–347. doi: 10.1126/science.806966. [DOI] [PubMed] [Google Scholar]
- Desplan C., Theis J., O'Farrell P. H. The Drosophila developmental gene, engrailed, encodes a sequence-specific DNA binding activity. Nature. 1985 Dec 19;318(6047):630–635. doi: 10.1038/318630a0. [DOI] [PMC free article] [PubMed] [Google Scholar]
- DiNardo S., Sher E., Heemskerk-Jongens J., Kassis J. A., O'Farrell P. H. Two-tiered regulation of spatially patterned engrailed gene expression during Drosophila embryogenesis. Nature. 1988 Apr 14;332(6165):604–609. doi: 10.1038/332604a0. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Ferguson E. L., Anderson K. V. Decapentaplegic acts as a morphogen to organize dorsal-ventral pattern in the Drosophila embryo. Cell. 1992 Oct 30;71(3):451–461. doi: 10.1016/0092-8674(92)90514-d. [DOI] [PubMed] [Google Scholar]
- Garcia-Bellido A., Santamaria P. Developmental analysis of the wing disc in the mutant engrailed of Drosophila melanogaster. Genetics. 1972 Sep;72(1):87–104. doi: 10.1093/genetics/72.1.87. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Gelbart W. M. Synapsis-dependent allelic complementation at the decapentaplegic gene complex in Drosophila melanogaster. Proc Natl Acad Sci U S A. 1982 Apr;79(8):2636–2640. doi: 10.1073/pnas.79.8.2636. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Gelbart W. M. The decapentaplegic gene: a TGF-beta homologue controlling pattern formation in Drosophila. Development. 1989;107 (Suppl):65–74. doi: 10.1242/dev.107.Supplement.65. [DOI] [PubMed] [Google Scholar]
- Golic K. G. Site-specific recombination between homologous chromosomes in Drosophila. Science. 1991 May 17;252(5008):958–961. doi: 10.1126/science.2035025. [DOI] [PubMed] [Google Scholar]
- Hoey T., Levine M. Divergent homeo box proteins recognize similar DNA sequences in Drosophila. Nature. 1988 Apr 28;332(6167):858–861. doi: 10.1038/332858a0. [DOI] [PubMed] [Google Scholar]
- Huang J. D., Schwyter D. H., Shirokawa J. M., Courey A. J. The interplay between multiple enhancer and silencer elements defines the pattern of decapentaplegic expression. Genes Dev. 1993 Apr;7(4):694–704. doi: 10.1101/gad.7.4.694. [DOI] [PubMed] [Google Scholar]
- Ingham P. W., Martinez Arias A. Boundaries and fields in early embryos. Cell. 1992 Jan 24;68(2):221–235. doi: 10.1016/0092-8674(92)90467-q. [DOI] [PubMed] [Google Scholar]
- Kingsley D. M. The TGF-beta superfamily: new members, new receptors, and new genetic tests of function in different organisms. Genes Dev. 1994 Jan;8(2):133–146. doi: 10.1101/gad.8.2.133. [DOI] [PubMed] [Google Scholar]
- Kornberg T., Sidén I., O'Farrell P., Simon M. The engrailed locus of Drosophila: in situ localization of transcripts reveals compartment-specific expression. Cell. 1985 Jan;40(1):45–53. doi: 10.1016/0092-8674(85)90307-1. [DOI] [PubMed] [Google Scholar]
- Lawrence P. A., Morata G. Compartments in the wing of Drosophila: a study of the engrailed gene. Dev Biol. 1976 Jun;50(2):321–337. doi: 10.1016/0012-1606(76)90155-x. [DOI] [PubMed] [Google Scholar]
- Lee J. J., von Kessler D. P., Parks S., Beachy P. A. Secretion and localized transcription suggest a role in positional signaling for products of the segmentation gene hedgehog. Cell. 1992 Oct 2;71(1):33–50. doi: 10.1016/0092-8674(92)90264-d. [DOI] [PubMed] [Google Scholar]
- Martinez-Arias A., Lawrence P. A. Parasegments and compartments in the Drosophila embryo. Nature. 1985 Feb 21;313(6004):639–642. doi: 10.1038/313639a0. [DOI] [PubMed] [Google Scholar]
- Masucci J. D., Miltenberger R. J., Hoffmann F. M. Pattern-specific expression of the Drosophila decapentaplegic gene in imaginal disks is regulated by 3' cis-regulatory elements. Genes Dev. 1990 Nov;4(11):2011–2023. doi: 10.1101/gad.4.11.2011. [DOI] [PubMed] [Google Scholar]
- Padgett R. W., St Johnston R. D., Gelbart W. M. A transcript from a Drosophila pattern gene predicts a protein homologous to the transforming growth factor-beta family. Nature. 1987 Jan 1;325(6099):81–84. doi: 10.1038/325081a0. [DOI] [PubMed] [Google Scholar]
- Perrimon N. The genetic basis of patterned baldness in Drosophila. Cell. 1994 Mar 11;76(5):781–784. doi: 10.1016/0092-8674(94)90351-4. [DOI] [PubMed] [Google Scholar]
- Phillips R. G., Roberts I. J., Ingham P. W., Whittle J. R. The Drosophila segment polarity gene patched is involved in a position-signalling mechanism in imaginal discs. Development. 1990 Sep;110(1):105–114. doi: 10.1242/dev.110.1.105. [DOI] [PubMed] [Google Scholar]
- Posakony L. G., Raftery L. A., Gelbart W. M. Wing formation in Drosophila melanogaster requires decapentaplegic gene function along the anterior-posterior compartment boundary. Mech Dev. 1990 Dec;33(1):69–82. doi: 10.1016/0925-4773(90)90136-a. [DOI] [PubMed] [Google Scholar]
- Raftery L. A., Sanicola M., Blackman R. K., Gelbart W. M. The relationship of decapentaplegic and engrailed expression in Drosophila imaginal disks: do these genes mark the anterior-posterior compartment boundary? Development. 1991 Sep;113(1):27–33. doi: 10.1242/dev.113.1.27. [DOI] [PubMed] [Google Scholar]
- Ray R. P., Arora K., Nüsslein-Volhard C., Gelbart W. M. The control of cell fate along the dorsal-ventral axis of the Drosophila embryo. Development. 1991 Sep;113(1):35–54. doi: 10.1242/dev.113.1.35. [DOI] [PubMed] [Google Scholar]
- Smith D. B., Johnson K. S. Single-step purification of polypeptides expressed in Escherichia coli as fusions with glutathione S-transferase. Gene. 1988 Jul 15;67(1):31–40. doi: 10.1016/0378-1119(88)90005-4. [DOI] [PubMed] [Google Scholar]
- Smolik-Utlaut S. M., Gelbart W. M. The effects of chromosomal rearrangements on the zeste-white interaction in Drosophila melanogaster. Genetics. 1987 Jun;116(2):285–298. doi: 10.1093/genetics/116.2.285. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Spencer F. A., Hoffmann F. M., Gelbart W. M. Decapentaplegic: a gene complex affecting morphogenesis in Drosophila melanogaster. Cell. 1982 Mar;28(3):451–461. doi: 10.1016/0092-8674(82)90199-4. [DOI] [PubMed] [Google Scholar]
- St Johnston D., Nüsslein-Volhard C. The origin of pattern and polarity in the Drosophila embryo. Cell. 1992 Jan 24;68(2):201–219. doi: 10.1016/0092-8674(92)90466-p. [DOI] [PubMed] [Google Scholar]
- St Johnston R. D., Hoffmann F. M., Blackman R. K., Segal D., Grimaila R., Padgett R. W., Irick H. A., Gelbart W. M. Molecular organization of the decapentaplegic gene in Drosophila melanogaster. Genes Dev. 1990 Jul;4(7):1114–1127. doi: 10.1101/gad.4.7.1114. [DOI] [PubMed] [Google Scholar]
- Tabata T., Eaton S., Kornberg T. B. The Drosophila hedgehog gene is expressed specifically in posterior compartment cells and is a target of engrailed regulation. Genes Dev. 1992 Dec;6(12B):2635–2645. doi: 10.1101/gad.6.12b.2635. [DOI] [PubMed] [Google Scholar]
- Tabata T., Kornberg T. B. Hedgehog is a signaling protein with a key role in patterning Drosophila imaginal discs. Cell. 1994 Jan 14;76(1):89–102. doi: 10.1016/0092-8674(94)90175-9. [DOI] [PubMed] [Google Scholar]
- Wharton K. A., Ray R. P., Gelbart W. M. An activity gradient of decapentaplegic is necessary for the specification of dorsal pattern elements in the Drosophila embryo. Development. 1993 Feb;117(2):807–822. doi: 10.1242/dev.117.2.807. [DOI] [PubMed] [Google Scholar]
- Williams J. A., Paddock S. W., Carroll S. B. Pattern formation in a secondary field: a hierarchy of regulatory genes subdivides the developing Drosophila wing disc into discrete subregions. Development. 1993 Feb;117(2):571–584. doi: 10.1242/dev.117.2.571. [DOI] [PubMed] [Google Scholar]