Skip to main content
NCBI home page
The Journal of Cell Biology logoLink to The Journal of Cell Biology
. 1989 Nov 1;109(5):2427–2440. doi: 10.1083/jcb.109.5.2427

The notch gene product is a glycoprotein expressed on the cell surface of both epidermal and neuronal precursor cells during Drosophila development

PMCID: PMC2115861  PMID: 2509485

Abstract

The Notch locus of Drosophila melanogaster is one of a small number of zygotically acting "neurogenic" genes involved in the correct segregation of neural from epidermal lineages during embryogenesis as well as in other postembryonic developmental events. We have generated antibody probes against three regions of the Notch protein to study the expression of Notch and begin a biochemical characterization of the protein. Consistent with predictions based on DNA sequence data, here we gather evidence showing that Notch encodes a large, glycosylated surface protein with an apparent molecular mass of 300 kD: (a) all three antibodies detect Notch on Western blots as a high molecular mass, primarily full-length product; (b) immunoelectron microscopy localizes the Notch protein to the cell membrane; and (c) lentil lectin column binding demonstrates that the protein is glycosylated, indicative of its surface protein nature. In general, the distribution of the Notch protein coincides with that of the Notch transcript determined previously by in situ hybridizations. Notch is expressed in a much wider range of tissue types than those disrupted in the neurogenic mutant, as determined by antibody localization. Early labeling in the blastoderm appears ubiquitous except for the pole cells, but as development proceeds some distinctive features emerge: stronger staining is seen within the germ band layer where neuroblast delamination occurs, and the developing embryonic nervous system shows pronounced axonal staining. In third instar larvae, Notch is expressed in imaginal disks and in the central nervous system. Based on these results, certain models for how Notch controls the neuroblast cell fate choice are eliminated. We discuss how Notch may function in this choice as well as in other lineage fate determinations.

Full Text

The Full Text of this article is available as a PDF (9.0 MB).

Selected References

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

  1. Artavanis-Tsakonas S., Muskavitch M. A., Yedvobnick B. Molecular cloning of Notch, a locus affecting neurogenesis in Drosophila melanogaster. Proc Natl Acad Sci U S A. 1983 Apr;80(7):1977–1981. doi: 10.1073/pnas.80.7.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Artavanis-Tsakonas S. The molecular biology of the Notch locus and the fine tuning of differentiation in Drosophila. Trends Genet. 1988 Apr;4(4):95–100. doi: 10.1016/0168-9525(88)90096-0. [DOI] [PubMed] [Google Scholar]
  3. Breeden L., Nasmyth K. Similarity between cell-cycle genes of budding yeast and fission yeast and the Notch gene of Drosophila. Nature. 1987 Oct 15;329(6140):651–654. doi: 10.1038/329651a0. [DOI] [PubMed] [Google Scholar]
  4. Dietrich U., Campos-Ortega J. A. The expression of neurogenic loci in imaginal epidermal cells of Drosophila melanogaster. J Neurogenet. 1984 Dec;1(4):315–332. doi: 10.3109/01677068409107094. [DOI] [PubMed] [Google Scholar]
  5. Doe C. Q., Goodman C. S. Early events in insect neurogenesis. I. Development and segmental differences in the pattern of neuronal precursor cells. Dev Biol. 1985 Sep;111(1):193–205. doi: 10.1016/0012-1606(85)90445-2. [DOI] [PubMed] [Google Scholar]
  6. Foster G. G. Negative complementation at the notch locus of Drosophila melanogaster. Genetics. 1975 Sep;81(1):99–120. doi: 10.1093/genetics/81.1.99. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Hartley D. A., Preiss A., Artavanis-Tsakonas S. A deduced gene product from the Drosophila neurogenic locus, enhancer of split, shows homology to mammalian G-protein beta subunit. Cell. 1988 Dec 2;55(5):785–795. doi: 10.1016/0092-8674(88)90134-1. [DOI] [PubMed] [Google Scholar]
  8. Hartley D. A., Xu T. A., Artavanis-Tsakonas S. The embryonic expression of the Notch locus of Drosophila melanogaster and the implications of point mutations in the extracellular EGF-like domain of the predicted protein. EMBO J. 1987 Nov;6(11):3407–3417. doi: 10.1002/j.1460-2075.1987.tb02664.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Hoppe P. E., Greenspan R. J. Local function of the Notch gene for embryonic ectodermal pathway choice in Drosophila. Cell. 1986 Aug 29;46(5):773–783. doi: 10.1016/0092-8674(86)90353-3. [DOI] [PubMed] [Google Scholar]
  10. Jessell T. M. Adhesion molecules and the hierarchy of neural development. Neuron. 1988 Mar;1(1):3–13. doi: 10.1016/0896-6273(88)90204-8. [DOI] [PubMed] [Google Scholar]
  11. Jones F. S., Burgoon M. P., Hoffman S., Crossin K. L., Cunningham B. A., Edelman G. M. A cDNA clone for cytotactin contains sequences similar to epidermal growth factor-like repeats and segments of fibronectin and fibrinogen. Proc Natl Acad Sci U S A. 1988 Apr;85(7):2186–2190. doi: 10.1073/pnas.85.7.2186. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Karr T. L., Kornberg T. B. fushi tarazu protein expression in the cellular blastoderm of Drosophila detected using a novel imaging technique. Development. 1989 May;106(1):95–103. doi: 10.1242/dev.106.1.95. [DOI] [PubMed] [Google Scholar]
  13. Kidd S., Lockett T. J., Young M. W. The Notch locus of Drosophila melanogaster. Cell. 1983 Sep;34(2):421–433. doi: 10.1016/0092-8674(83)90376-8. [DOI] [PubMed] [Google Scholar]
  14. Knust E., Dietrich U., Tepass U., Bremer K. A., Weigel D., Vässin H., Campos-Ortega J. A. EGF homologous sequences encoded in the genome of Drosophila melanogaster, and their relation to neurogenic genes. EMBO J. 1987 Mar;6(3):761–766. doi: 10.1002/j.1460-2075.1987.tb04818.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Lefevre G., Jr, Green M. M. Genetic duplication in the white-split interval of the X chromosome in Drosophila melanogaster. Chromosoma. 1972;36(4):391–412. doi: 10.1007/BF00336795. [DOI] [PubMed] [Google Scholar]
  16. Liposits Z., Sétáló G., Flerkó B. Application of the silver-gold intensified 3,3'-diaminobenzidine chromogen to the light and electron microscopic detection of the luteinizing hormone-releasing hormone system of the rat brain. Neuroscience. 1984 Oct;13(2):513–525. doi: 10.1016/0306-4522(84)90245-8. [DOI] [PubMed] [Google Scholar]
  17. Markopoulou K., Artavanis-Tsakonas S. The expression of the neurogenic locus Notch during the postembryonic development of Drosophila melanogaster and its relationship to mitotic activity. J Neurogenet. 1989 Sep;6(1):11–26. doi: 10.3109/01677068909107097. [DOI] [PubMed] [Google Scholar]
  18. McCormick F., Clark B. F., la Cour T. F., Kjeldgaard M., Norskov-Lauritsen L., Nyborg J. A model for the tertiary structure of p21, the product of the ras oncogene. Science. 1985 Oct 4;230(4721):78–82. doi: 10.1126/science.3898366. [DOI] [PubMed] [Google Scholar]
  19. Mitchison T. J., Sedat J. Localization of antigenic determinants in whole Drosophila embryos. Dev Biol. 1983 Sep;99(1):261–264. doi: 10.1016/0012-1606(83)90275-0. [DOI] [PubMed] [Google Scholar]
  20. Perrimon N., Engstrom L., Mahowald A. P. A pupal lethal mutation with a paternally influenced maternal effect on embryonic development in Drosophila melanogaster. Dev Biol. 1985 Aug;110(2):480–491. doi: 10.1016/0012-1606(85)90105-8. [DOI] [PubMed] [Google Scholar]
  21. Portin P. Allelic negative complementation at the Abruptex locus of Drosophila melanogaster. Genetics. 1975 Sep;81(1):121–133. doi: 10.1093/genetics/81.1.121. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Poulson D. F. Chromosomal Deficiencies and the Embryonic Development of Drosophila Melanogaster. Proc Natl Acad Sci U S A. 1937 Mar;23(3):133–137. doi: 10.1073/pnas.23.3.133. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Preiss A., Hartley D. A., Artavanis-Tsakonas S. The molecular genetics of Enhancer of split, a gene required for embryonic neural development in Drosophila. EMBO J. 1988 Dec 1;7(12):3917–3927. doi: 10.1002/j.1460-2075.1988.tb03278.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Rothberg J. M., Hartley D. A., Walther Z., Artavanis-Tsakonas S. slit: an EGF-homologous locus of D. melanogaster involved in the development of the embryonic central nervous system. Cell. 1988 Dec 23;55(6):1047–1059. doi: 10.1016/0092-8674(88)90249-8. [DOI] [PubMed] [Google Scholar]
  25. Rüther U., Müller-Hill B. Easy identification of cDNA clones. EMBO J. 1983;2(10):1791–1794. doi: 10.1002/j.1460-2075.1983.tb01659.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Schubiger M., Palka J. Changing spatial patterns of DNA replication in the developing wing of Drosophila. Dev Biol. 1987 Sep;123(1):145–153. doi: 10.1016/0012-1606(87)90436-2. [DOI] [PubMed] [Google Scholar]
  27. Schüpbach T., Wieschaus E. Female sterile mutations on the second chromosome of Drosophila melanogaster. I. Maternal effect mutations. Genetics. 1989 Jan;121(1):101–117. doi: 10.1093/genetics/121.1.101. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Shellenbarger D. L., Mohler J. D. Temperature-sensitive periods and autonomy of pleiotropic effects of l(1)Nts1, a conditional notch lethal in Drosophila. Dev Biol. 1978 Feb;62(2):432–446. doi: 10.1016/0012-1606(78)90226-9. [DOI] [PubMed] [Google Scholar]
  29. Stern C., Tokunaga C. Autonomous pleiotropy in Drosophilia. Proc Natl Acad Sci U S A. 1968 Aug;60(4):1252–1259. doi: 10.1073/pnas.60.4.1252. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Tan S. S., Crossin K. L., Hoffman S., Edelman G. M. Asymmetric expression in somites of cytotactin and its proteoglycan ligand is correlated with neural crest cell distribution. Proc Natl Acad Sci U S A. 1987 Nov;84(22):7977–7981. doi: 10.1073/pnas.84.22.7977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Technau G. M., Campos-Ortega J. A. Cell autonomy of expression of neurogenic genes of Drosophila melanogaster. Proc Natl Acad Sci U S A. 1987 Jul;84(13):4500–4504. doi: 10.1073/pnas.84.13.4500. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Towbin H., Staehelin T., Gordon J. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4350–4354. doi: 10.1073/pnas.76.9.4350. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Truman J. W., Bate M. Spatial and temporal patterns of neurogenesis in the central nervous system of Drosophila melanogaster. Dev Biol. 1988 Jan;125(1):145–157. doi: 10.1016/0012-1606(88)90067-x. [DOI] [PubMed] [Google Scholar]
  34. Vässin H., Bremer K. A., Knust E., Campos-Ortega J. A. The neurogenic gene Delta of Drosophila melanogaster is expressed in neurogenic territories and encodes a putative transmembrane protein with EGF-like repeats. EMBO J. 1987 Nov;6(11):3431–3440. doi: 10.1002/j.1460-2075.1987.tb02666.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. WELSHONS W. J., VON HALLE E. S. Pseudoallelism at the notch locus in drosophila. Genetics. 1962 Jun;47:743–759. doi: 10.1093/genetics/47.6.743. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Welshons W. J. Analysis of a gene in drosophila. Science. 1965 Nov 26;150(3700):1122–1129. doi: 10.1126/science.150.3700.1122. [DOI] [PubMed] [Google Scholar]
  37. Welshons W. J. Genetic basis for two types of recessive lethality at the notch locus of Drosophila. Genetics. 1971 Jun;68(2):259–268. doi: 10.1093/genetics/68.2.259. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Welshons W. J., Keppy D. O. The recombinational analysis of aberrations and the position of the notch locus on the polytene chromosome of Drosophila. Mol Gen Genet. 1981;181(3):319–324. doi: 10.1007/BF00425605. [DOI] [PubMed] [Google Scholar]
  39. Wharton K. A., Johansen K. M., Xu T., Artavanis-Tsakonas S. Nucleotide sequence from the neurogenic locus notch implies a gene product that shares homology with proteins containing EGF-like repeats. Cell. 1985 Dec;43(3 Pt 2):567–581. doi: 10.1016/0092-8674(85)90229-6. [DOI] [PubMed] [Google Scholar]
  40. Wharton K. A., Yedvobnick B., Finnerty V. G., Artavanis-Tsakonas S. opa: a novel family of transcribed repeats shared by the Notch locus and other developmentally regulated loci in D. melanogaster. Cell. 1985 Jan;40(1):55–62. doi: 10.1016/0092-8674(85)90308-3. [DOI] [PubMed] [Google Scholar]
  41. White K., Kankel D. R. Patterns of cell division and cell movement in the formation of the imaginal nervous system in Drosophila melanogaster. Dev Biol. 1978 Aug;65(2):296–321. doi: 10.1016/0012-1606(78)90029-5. [DOI] [PubMed] [Google Scholar]
  42. Wright T. R. The genetics of embryogenesis in Drosophila. Adv Genet. 1970;15:261–395. doi: 10.1016/s0065-2660(08)60075-9. [DOI] [PubMed] [Google Scholar]
  43. Yedvobnick B., Muskavitch M. A., Wharton K. A., Halpern M. E., Paul E., Grimwade B. G., Artavanis-Tsakonas S. Molecular genetics of Drosophila neurogenesis. Cold Spring Harb Symp Quant Biol. 1985;50:841–854. doi: 10.1101/sqb.1985.050.01.102. [DOI] [PubMed] [Google Scholar]
  44. Yochem J., Weston K., Greenwald I. The Caenorhabditis elegans lin-12 gene encodes a transmembrane protein with overall similarity to Drosophila Notch. Nature. 1988 Oct 6;335(6190):547–550. doi: 10.1038/335547a0. [DOI] [PubMed] [Google Scholar]

Articles from The Journal of Cell Biology are provided here courtesy of The Rockefeller University Press

RESOURCES