Skip to main content
NCBI home page
Genetics logoLink to Genetics
. 1996 May;143(1):315–329. doi: 10.1093/genetics/143.1.315

A Screen for Genes That Function Downstream of Ras1 during Drosophila Eye Development

F D Karim 1, H C Chang 1, M Therrien 1, D A Wassarman 1, T Laverty 1, G M Rubin 1
PMCID: PMC1207264  PMID: 8722784

Abstract

Cell-fate specification of the R7 photoreceptor cell is controlled by the sevenless receptor tyrosine kinase (SevRTK) and Ras1, the Drosophila homologue of mammalian H-ras, K-ras and N-ras oncogenes. An activated form of Ras1 expressed under control of the sevenless enhancer/promoter (sev-Ras1(V12)) induces production of supernumerary R7 photoreceptor cells, which causes the eye to become rough in appearance. To isolate mutations in genes functioning downstream of Ras1, we carried out a screen for dominant suppressors and enhancers of this rough eye phenotype. Approximately 850,000 mutagenized flies were screened, and 282 dominant suppressors and 577 dominant enhancers were isolated. Mutations in the Drosophila homologues of Raf, MEK, MAPK, type I Geranylgeranyl Transferase and Protein Phosphatase 2A were isolated, as were mutations in several novel signaling genes. Some of these mutant genes appear to be general signaling factors that function in other Ras1 pathways, while one seems to be more specific for photoreceptor development. At least two suppressors appear to function either between Ras1 and Raf or in parallele to Raf.

Full Text

The Full Text of this article is available as a PDF (6.6 MB).

Selected References

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

  1. Ambrosio L., Mahowald A. P., Perrimon N. Requirement of the Drosophila raf homologue for torso function. Nature. 1989 Nov 16;342(6247):288–291. doi: 10.1038/342288a0. [DOI] [PubMed] [Google Scholar]
  2. Basler K., Christen B., Hafen E. Ligand-independent activation of the sevenless receptor tyrosine kinase changes the fate of cells in the developing Drosophila eye. Cell. 1991 Mar 22;64(6):1069–1081. doi: 10.1016/0092-8674(91)90262-w. [DOI] [PubMed] [Google Scholar]
  3. Biggs W. H., 3rd, Zavitz K. H., Dickson B., van der Straten A., Brunner D., Hafen E., Zipursky S. L. The Drosophila rolled locus encodes a MAP kinase required in the sevenless signal transduction pathway. EMBO J. 1994 Apr 1;13(7):1628–1635. doi: 10.1002/j.1460-2075.1994.tb06426.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Biggs W. H., 3rd, Zipursky S. L. Primary structure, expression, and signal-dependent tyrosine phosphorylation of a Drosophila homolog of extracellular signal-regulated kinase. Proc Natl Acad Sci U S A. 1992 Jul 15;89(14):6295–6299. doi: 10.1073/pnas.89.14.6295. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Bohmann D., Ellis M. C., Staszewski L. M., Mlodzik M. Drosophila Jun mediates Ras-dependent photoreceptor determination. Cell. 1994 Sep 23;78(6):973–986. doi: 10.1016/0092-8674(94)90273-9. [DOI] [PubMed] [Google Scholar]
  6. Bruder J. T., Heidecker G., Rapp U. R. Serum-, TPA-, and Ras-induced expression from Ap-1/Ets-driven promoters requires Raf-1 kinase. Genes Dev. 1992 Apr;6(4):545–556. doi: 10.1101/gad.6.4.545. [DOI] [PubMed] [Google Scholar]
  7. Buss J. E., Solski P. A., Schaeffer J. P., MacDonald M. J., Der C. J. Activation of the cellular proto-oncogene product p21Ras by addition of a myristylation signal. Science. 1989 Mar 24;243(4898):1600–1603. doi: 10.1126/science.2648572. [DOI] [PubMed] [Google Scholar]
  8. Carthew R. W., Rubin G. M. seven in absentia, a gene required for specification of R7 cell fate in the Drosophila eye. Cell. 1990 Nov 2;63(3):561–577. doi: 10.1016/0092-8674(90)90452-k. [DOI] [PubMed] [Google Scholar]
  9. Chang H. C., Solomon N. M., Wassarman D. A., Karim F. D., Therrien M., Rubin G. M., Wolff T. phyllopod functions in the fate determination of a subset of photoreceptors in Drosophila. Cell. 1995 Feb 10;80(3):463–472. doi: 10.1016/0092-8674(95)90497-2. [DOI] [PubMed] [Google Scholar]
  10. Diaz-Benjumea F. J., Hafen E. The sevenless signalling cassette mediates Drosophila EGF receptor function during epidermal development. Development. 1994 Mar;120(3):569–578. doi: 10.1242/dev.120.3.569. [DOI] [PubMed] [Google Scholar]
  11. Dickson B. J., Domínguez M., van der Straten A., Hafen E. Control of Drosophila photoreceptor cell fates by phyllopod, a novel nuclear protein acting downstream of the Raf kinase. Cell. 1995 Feb 10;80(3):453–462. doi: 10.1016/0092-8674(95)90496-4. [DOI] [PubMed] [Google Scholar]
  12. Duffy J. B., Perrimon N. The torso pathway in Drosophila: lessons on receptor tyrosine kinase signaling and pattern formation. Dev Biol. 1994 Dec;166(2):380–395. doi: 10.1006/dbio.1994.1324. [DOI] [PubMed] [Google Scholar]
  13. Feig L. A., Cooper G. M. Inhibition of NIH 3T3 cell proliferation by a mutant ras protein with preferential affinity for GDP. Mol Cell Biol. 1988 Aug;8(8):3235–3243. doi: 10.1128/mcb.8.8.3235. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Fortini M. E., Simon M. A., Rubin G. M. Signalling by the sevenless protein tyrosine kinase is mimicked by Ras1 activation. Nature. 1992 Feb 6;355(6360):559–561. doi: 10.1038/355559a0. [DOI] [PubMed] [Google Scholar]
  15. Gaul U., Mardon G., Rubin G. M. A putative Ras GTPase activating protein acts as a negative regulator of signaling by the Sevenless receptor tyrosine kinase. Cell. 1992 Mar 20;68(6):1007–1019. doi: 10.1016/0092-8674(92)90073-l. [DOI] [PubMed] [Google Scholar]
  16. Hoey T., Weinzierl R. O., Gill G., Chen J. L., Dynlacht B. D., Tjian R. Molecular cloning and functional analysis of Drosophila TAF110 reveal properties expected of coactivators. Cell. 1993 Jan 29;72(2):247–260. doi: 10.1016/0092-8674(93)90664-c. [DOI] [PubMed] [Google Scholar]
  17. Kaplan J. M., Mardon G., Bishop J. M., Varmus H. E. The first seven amino acids encoded by the v-src oncogene act as a myristylation signal: lysine 7 is a critical determinant. Mol Cell Biol. 1988 Jun;8(6):2435–2441. doi: 10.1128/mcb.8.6.2435. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Karpen G. H., Spradling A. C. Analysis of subtelomeric heterochromatin in the Drosophila minichromosome Dp1187 by single P element insertional mutagenesis. Genetics. 1992 Nov;132(3):737–753. doi: 10.1093/genetics/132.3.737. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Kimmel B. E., Heberlein U., Rubin G. M. The homeo domain protein rough is expressed in a subset of cells in the developing Drosophila eye where it can specify photoreceptor cell subtype. Genes Dev. 1990 May;4(5):712–727. doi: 10.1101/gad.4.5.712. [DOI] [PubMed] [Google Scholar]
  20. Lai Z. C., Rubin G. M. Negative control of photoreceptor development in Drosophila by the product of the yan gene, an ETS domain protein. Cell. 1992 Aug 21;70(4):609–620. doi: 10.1016/0092-8674(92)90430-k. [DOI] [PubMed] [Google Scholar]
  21. Lowy D. R., Willumsen B. M. Function and regulation of ras. Annu Rev Biochem. 1993;62:851–891. doi: 10.1146/annurev.bi.62.070193.004223. [DOI] [PubMed] [Google Scholar]
  22. Melnick M. B., Perkins L. A., Lee M., Ambrosio L., Perrimon N. Developmental and molecular characterization of mutations in the Drosophila-raf serine/threonine protein kinase. Development. 1993 May;118(1):127–138. doi: 10.1242/dev.118.1.127. [DOI] [PubMed] [Google Scholar]
  23. Moodie S. A., Wolfman A. The 3Rs of life: Ras, Raf and growth regulation. Trends Genet. 1994 Feb;10(2):44–48. doi: 10.1016/0168-9525(94)90147-3. [DOI] [PubMed] [Google Scholar]
  24. Muskavitch M. A. Delta-notch signaling and Drosophila cell fate choice. Dev Biol. 1994 Dec;166(2):415–430. doi: 10.1006/dbio.1994.1326. [DOI] [PubMed] [Google Scholar]
  25. Nishida Y., Hata M., Ayaki T., Ryo H., Yamagata M., Shimizu K., Nishizuka Y. Proliferation of both somatic and germ cells is affected in the Drosophila mutants of raf proto-oncogene. EMBO J. 1988 Mar;7(3):775–781. doi: 10.1002/j.1460-2075.1988.tb02875.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. O'Neill E. M., Rebay I., Tjian R., Rubin G. M. The activities of two Ets-related transcription factors required for Drosophila eye development are modulated by the Ras/MAPK pathway. Cell. 1994 Jul 15;78(1):137–147. doi: 10.1016/0092-8674(94)90580-0. [DOI] [PubMed] [Google Scholar]
  27. Olivier J. P., Raabe T., Henkemeyer M., Dickson B., Mbamalu G., Margolis B., Schlessinger J., Hafen E., Pawson T. A Drosophila SH2-SH3 adaptor protein implicated in coupling the sevenless tyrosine kinase to an activator of Ras guanine nucleotide exchange, Sos. Cell. 1993 Apr 9;73(1):179–191. doi: 10.1016/0092-8674(93)90170-u. [DOI] [PubMed] [Google Scholar]
  28. Rebay I., Rubin G. M. Yan functions as a general inhibitor of differentiation and is negatively regulated by activation of the Ras1/MAPK pathway. Cell. 1995 Jun 16;81(6):857–866. doi: 10.1016/0092-8674(95)90006-3. [DOI] [PubMed] [Google Scholar]
  29. Schlessinger J. How receptor tyrosine kinases activate Ras. Trends Biochem Sci. 1993 Aug;18(8):273–275. doi: 10.1016/0968-0004(93)90031-h. [DOI] [PubMed] [Google Scholar]
  30. Sigal I. S., Gibbs J. B., D'Alonzo J. S., Temeles G. L., Wolanski B. S., Socher S. H., Scolnick E. M. Mutant ras-encoded proteins with altered nucleotide binding exert dominant biological effects. Proc Natl Acad Sci U S A. 1986 Feb;83(4):952–956. doi: 10.1073/pnas.83.4.952. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Simon M. A., Dodson G. S., Rubin G. M. An SH3-SH2-SH3 protein is required for p21Ras1 activation and binds to sevenless and Sos proteins in vitro. Cell. 1993 Apr 9;73(1):169–177. doi: 10.1016/0092-8674(93)90169-q. [DOI] [PubMed] [Google Scholar]
  32. Simon M. A. Signal transduction during the development of the Drosophila R7 photoreceptor. Dev Biol. 1994 Dec;166(2):431–442. doi: 10.1006/dbio.1994.1327. [DOI] [PubMed] [Google Scholar]
  33. Therrien M., Chang H. C., Solomon N. M., Karim F. D., Wassarman D. A., Rubin G. M. KSR, a novel protein kinase required for RAS signal transduction. Cell. 1995 Dec 15;83(6):879–888. doi: 10.1016/0092-8674(95)90204-x. [DOI] [PubMed] [Google Scholar]
  34. Tomlinson A., Ready D. F. Neuronal differentiation in Drosophila ommatidium. Dev Biol. 1987 Apr;120(2):366–376. doi: 10.1016/0012-1606(87)90239-9. [DOI] [PubMed] [Google Scholar]
  35. Tsuda L., Inoue Y. H., Yoo M. A., Mizuno M., Hata M., Lim Y. M., Adachi-Yamada T., Ryo H., Masamune Y., Nishida Y. A protein kinase similar to MAP kinase activator acts downstream of the raf kinase in Drosophila. Cell. 1993 Feb 12;72(3):407–414. doi: 10.1016/0092-8674(93)90117-9. [DOI] [PubMed] [Google Scholar]
  36. Török T., Tick G., Alvarado M., Kiss I. P-lacW insertional mutagenesis on the second chromosome of Drosophila melanogaster: isolation of lethals with different overgrowth phenotypes. Genetics. 1993 Sep;135(1):71–80. doi: 10.1093/genetics/135.1.71. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Van Vactor D. L., Jr, Cagan R. L., Krämer H., Zipursky S. L. Induction in the developing compound eye of Drosophila: multiple mechanisms restrict R7 induction to a single retinal precursor cell. Cell. 1991 Dec 20;67(6):1145–1155. doi: 10.1016/0092-8674(91)90291-6. [DOI] [PubMed] [Google Scholar]
  38. Wassarman D. A., Solomon N. M., Chang H. C., Karim F. D., Therrien M., Rubin G. M. Protein phosphatase 2A positively and negatively regulates Ras1-mediated photoreceptor development in Drosophila. Genes Dev. 1996 Feb 1;10(3):272–278. doi: 10.1101/gad.10.3.272. [DOI] [PubMed] [Google Scholar]
  39. Wassarman D. A., Therrien M., Rubin G. M. The Ras signaling pathway in Drosophila. Curr Opin Genet Dev. 1995 Feb;5(1):44–50. doi: 10.1016/s0959-437x(95)90052-7. [DOI] [PubMed] [Google Scholar]
  40. Zipursky S. L., Rubin G. M. Determination of neuronal cell fate: lessons from the R7 neuron of Drosophila. Annu Rev Neurosci. 1994;17:373–397. doi: 10.1146/annurev.ne.17.030194.002105. [DOI] [PubMed] [Google Scholar]
  41. van der Geer P., Hunter T., Lindberg R. A. Receptor protein-tyrosine kinases and their signal transduction pathways. Annu Rev Cell Biol. 1994;10:251–337. doi: 10.1146/annurev.cb.10.110194.001343. [DOI] [PubMed] [Google Scholar]

Articles from Genetics are provided here courtesy of Oxford University Press

RESOURCES