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. 2000 Nov;156(3):1219–1230. doi: 10.1093/genetics/156.3.1219

A misexpression screen identifies genes that can modulate RAS1 pathway signaling in Drosophila melanogaster.

A M Huang 1, G M Rubin 1
PMCID: PMC1461302  PMID: 11063696

Abstract

Differentiation of the R7 photoreceptor cell is dependent on the Sevenless receptor tyrosine kinase, which activates the RAS1/mitogen-activated protein kinase signaling cascade. Kinase suppressor of Ras (KSR) functions genetically downstream of RAS1 in this signal transduction cascade. Expression of dominant-negative KSR (KDN) in the developing eye blocks RAS pathway signaling, prevents R7 cell differentiation, and causes a rough eye phenotype. To identify genes that modulate RAS signaling, we screened for genes that alter RAS1/KSR signaling efficiency when misexpressed. In this screen, we recovered three known genes, Lk6, misshapen, and Akap200. We also identified seven previously undescribed genes; one encodes a novel rel domain member of the NFAT family, and six encode novel proteins. These genes may represent new components of the RAS pathway or components of other signaling pathways that can modulate signaling by RAS. We discuss the utility of gain-of-function screens in identifying new components of signaling pathways in Drosophila.

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Selected References

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  1. Asha H., de Ruiter N. D., Wang M. G., Hariharan I. K. The Rap1 GTPase functions as a regulator of morphogenesis in vivo. EMBO J. 1999 Feb 1;18(3):605–615. doi: 10.1093/emboj/18.3.605. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Ashburner M., Misra S., Roote J., Lewis S. E., Blazej R., Davis T., Doyle C., Galle R., George R., Harris N. An exploration of the sequence of a 2.9-Mb region of the genome of Drosophila melanogaster: the Adh region. Genetics. 1999 Sep;153(1):179–219. doi: 10.1093/genetics/153.1.179. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bender A., Pringle J. R. Multicopy suppression of the cdc24 budding defect in yeast by CDC42 and three newly identified genes including the ras-related gene RSR1. Proc Natl Acad Sci U S A. 1989 Dec;86(24):9976–9980. doi: 10.1073/pnas.86.24.9976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Bollag G., McCormick F. Regulators and effectors of ras proteins. Annu Rev Cell Biol. 1991;7:601–632. doi: 10.1146/annurev.cb.07.110191.003125. [DOI] [PubMed] [Google Scholar]
  5. Brand A. H., Manoukian A. S., Perrimon N. Ectopic expression in Drosophila. Methods Cell Biol. 1994;44:635–654. doi: 10.1016/s0091-679x(08)60936-x. [DOI] [PubMed] [Google Scholar]
  6. Brand A. H., Perrimon N. Targeted gene expression as a means of altering cell fates and generating dominant phenotypes. Development. 1993 Jun;118(2):401–415. doi: 10.1242/dev.118.2.401. [DOI] [PubMed] [Google Scholar]
  7. Chen L., Glover J. N., Hogan P. G., Rao A., Harrison S. C. Structure of the DNA-binding domains from NFAT, Fos and Jun bound specifically to DNA. Nature. 1998 Mar 5;392(6671):42–48. doi: 10.1038/32100. [DOI] [PubMed] [Google Scholar]
  8. Colledge M., Scott J. D. AKAPs: from structure to function. Trends Cell Biol. 1999 Jun;9(6):216–221. doi: 10.1016/s0962-8924(99)01558-5. [DOI] [PubMed] [Google Scholar]
  9. Davis R. L., Weintraub H., Lassar A. B. Expression of a single transfected cDNA converts fibroblasts to myoblasts. Cell. 1987 Dec 24;51(6):987–1000. doi: 10.1016/0092-8674(87)90585-x. [DOI] [PubMed] [Google Scholar]
  10. Denko N. C., Giaccia A. J., Stringer J. R., Stambrook P. J. The human Ha-ras oncogene induces genomic instability in murine fibroblasts within one cell cycle. Proc Natl Acad Sci U S A. 1994 May 24;91(11):5124–5128. doi: 10.1073/pnas.91.11.5124. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Denouel-Galy A., Douville E. M., Warne P. H., Papin C., Laugier D., Calothy G., Downward J., Eychène A. Murine Ksr interacts with MEK and inhibits Ras-induced transformation. Curr Biol. 1998 Jan 1;8(1):46–55. doi: 10.1016/s0960-9822(98)70019-3. [DOI] [PubMed] [Google Scholar]
  12. Dickson B. J., van der Straten A., Dominguez M., Hafen E. Mutations Modulating Raf signaling in Drosophila eye development. Genetics. 1996 Jan;142(1):163–171. doi: 10.1093/genetics/142.1.163. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Dushay M. S., Asling B., Hultmark D. Origins of immunity: Relish, a compound Rel-like gene in the antibacterial defense of Drosophila. Proc Natl Acad Sci U S A. 1996 Sep 17;93(19):10343–10347. doi: 10.1073/pnas.93.19.10343. [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. Herskowitz I. MAP kinase pathways in yeast: for mating and more. Cell. 1995 Jan 27;80(2):187–197. doi: 10.1016/0092-8674(95)90402-6. [DOI] [PubMed] [Google Scholar]
  16. Ip Y. T., Reach M., Engstrom Y., Kadalayil L., Cai H., González-Crespo S., Tatei K., Levine M. Dif, a dorsal-related gene that mediates an immune response in Drosophila. Cell. 1993 Nov 19;75(4):753–763. doi: 10.1016/0092-8674(93)90495-c. [DOI] [PubMed] [Google Scholar]
  17. Jain J., Burgeon E., Badalian T. M., Hogan P. G., Rao A. A similar DNA-binding motif in NFAT family proteins and the Rel homology region. J Biol Chem. 1995 Feb 24;270(8):4138–4145. [PubMed] [Google Scholar]
  18. Jiang J., Struhl G. Protein kinase A and hedgehog signaling in Drosophila limb development. Cell. 1995 Feb 24;80(4):563–572. doi: 10.1016/0092-8674(95)90510-3. [DOI] [PubMed] [Google Scholar]
  19. Kitayama H., Sugimoto Y., Matsuzaki T., Ikawa Y., Noda M. A ras-related gene with transformation suppressor activity. Cell. 1989 Jan 13;56(1):77–84. doi: 10.1016/0092-8674(89)90985-9. [DOI] [PubMed] [Google Scholar]
  20. Kockel L., Zeitlinger J., Staszewski L. M., Mlodzik M., Bohmann D. Jun in Drosophila development: redundant and nonredundant functions and regulation by two MAPK signal transduction pathways. Genes Dev. 1997 Jul 1;11(13):1748–1758. doi: 10.1101/gad.11.13.1748. [DOI] [PubMed] [Google Scholar]
  21. Kornfeld K., Hom D. B., Horvitz H. R. The ksr-1 gene encodes a novel protein kinase involved in Ras-mediated signaling in C. elegans. Cell. 1995 Dec 15;83(6):903–913. doi: 10.1016/0092-8674(95)90206-6. [DOI] [PubMed] [Google Scholar]
  22. Li Z., Rossi E. A., Hoheisel J. D., Kalderon D., Rubin C. S. Generation of a novel A kinase anchor protein and a myristoylated alanine-rich C kinase substrate-like analog from a single gene. J Biol Chem. 1999 Sep 17;274(38):27191–27200. doi: 10.1074/jbc.274.38.27191. [DOI] [PubMed] [Google Scholar]
  23. Liao G. C., Rehm E. J., Rubin G. M. Insertion site preferences of the P transposable element in Drosophila melanogaster. Proc Natl Acad Sci U S A. 2000 Mar 28;97(7):3347–3351. doi: 10.1073/pnas.050017397. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Lopez-Rodríguez C., Aramburu J., Rakeman A. S., Rao A. NFAT5, a constitutively nuclear NFAT protein that does not cooperate with Fos and Jun. Proc Natl Acad Sci U S A. 1999 Jun 22;96(13):7214–7219. doi: 10.1073/pnas.96.13.7214. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Maixner A., Hecker T. P., Phan Q. N., Wassarman D. A. A screen for mutations that prevent lethality caused by expression of activated sevenless and Ras1 in the Drosophila embryo. Dev Genet. 1998;23(4):347–361. doi: 10.1002/(SICI)1520-6408(1998)23:4<347::AID-DVG9>3.0.CO;2-C. [DOI] [PubMed] [Google Scholar]
  26. Milán M., Diaz-Benjumea F. J., Cohen S. M. Beadex encodes an LMO protein that regulates Apterous LIM-homeodomain activity in Drosophila wing development: a model for LMO oncogene function. Genes Dev. 1998 Sep 15;12(18):2912–2920. doi: 10.1101/gad.12.18.2912. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Neufeld T. P., Tang A. H., Rubin G. M. A genetic screen to identify components of the sina signaling pathway in Drosophila eye development. Genetics. 1998 Jan;148(1):277–286. doi: 10.1093/genetics/148.1.277. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Pan D., Rubin G. M. cAMP-dependent protein kinase and hedgehog act antagonistically in regulating decapentaplegic transcription in Drosophila imaginal discs. Cell. 1995 Feb 24;80(4):543–552. doi: 10.1016/0092-8674(95)90508-1. [DOI] [PubMed] [Google Scholar]
  29. Paricio N., Feiguin F., Boutros M., Eaton S., Mlodzik M. The Drosophila STE20-like kinase misshapen is required downstream of the Frizzled receptor in planar polarity signaling. EMBO J. 1999 Sep 1;18(17):4669–4678. doi: 10.1093/emboj/18.17.4669. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Ramer S. W., Elledge S. J., Davis R. W. Dominant genetics using a yeast genomic library under the control of a strong inducible promoter. Proc Natl Acad Sci U S A. 1992 Dec 1;89(23):11589–11593. doi: 10.1073/pnas.89.23.11589. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Rao A., Luo C., Hogan P. G. Transcription factors of the NFAT family: regulation and function. Annu Rev Immunol. 1997;15:707–747. doi: 10.1146/annurev.immunol.15.1.707. [DOI] [PubMed] [Google Scholar]
  32. Rebay I., Chen F., Hsiao F., Kolodziej P. A., Kuang B. H., Laverty T., Suh C., Voas M., Williams A., Rubin G. M. A genetic screen for novel components of the Ras/Mitogen-activated protein kinase signaling pathway that interact with the yan gene of Drosophila identifies split ends, a new RNA recognition motif-containing protein. Genetics. 2000 Feb;154(2):695–712. doi: 10.1093/genetics/154.2.695. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Rine J., Hansen W., Hardeman E., Davis R. W. Targeted selection of recombinant clones through gene dosage effects. Proc Natl Acad Sci U S A. 1983 Nov;80(22):6750–6754. doi: 10.1073/pnas.80.22.6750. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Rørth P., Szabo K., Bailey A., Laverty T., Rehm J., Rubin G. M., Weigmann K., Milán M., Benes V., Ansorge W. Systematic gain-of-function genetics in Drosophila. Development. 1998 Mar;125(6):1049–1057. doi: 10.1242/dev.125.6.1049. [DOI] [PubMed] [Google Scholar]
  35. Saavedra H. I., Fukasawa K., Conn C. W., Stambrook P. J. MAPK mediates RAS-induced chromosome instability. J Biol Chem. 1999 Dec 31;274(53):38083–38090. doi: 10.1074/jbc.274.53.38083. [DOI] [PubMed] [Google Scholar]
  36. Seger R., Krebs E. G. The MAPK signaling cascade. FASEB J. 1995 Jun;9(9):726–735. [PubMed] [Google Scholar]
  37. Simon M. A., Bowtell D. D., Dodson G. S., Laverty T. R., Rubin G. M. Ras1 and a putative guanine nucleotide exchange factor perform crucial steps in signaling by the sevenless protein tyrosine kinase. Cell. 1991 Nov 15;67(4):701–716. doi: 10.1016/0092-8674(91)90065-7. [DOI] [PubMed] [Google Scholar]
  38. Smith W. C., Harland R. M. Expression cloning of noggin, a new dorsalizing factor localized to the Spemann organizer in Xenopus embryos. Cell. 1992 Sep 4;70(5):829–840. doi: 10.1016/0092-8674(92)90316-5. [DOI] [PubMed] [Google Scholar]
  39. 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]
  40. Therrien M., Michaud N. R., Rubin G. M., Morrison D. K. KSR modulates signal propagation within the MAPK cascade. Genes Dev. 1996 Nov 1;10(21):2684–2695. doi: 10.1101/gad.10.21.2684. [DOI] [PubMed] [Google Scholar]
  41. Therrien M., Morrison D. K., Wong A. M., Rubin G. M. A genetic screen for modifiers of a kinase suppressor of Ras-dependent rough eye phenotype in Drosophila. Genetics. 2000 Nov;156(3):1231–1242. doi: 10.1093/genetics/156.3.1231. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Therrien M., Wong A. M., Rubin G. M. CNK, a RAF-binding multidomain protein required for RAS signaling. Cell. 1998 Oct 30;95(3):343–353. doi: 10.1016/s0092-8674(00)81766-3. [DOI] [PubMed] [Google Scholar]
  43. Toba G., Ohsako T., Miyata N., Ohtsuka T., Seong K. H., Aigaki T. The gene search system. A method for efficient detection and rapid molecular identification of genes in Drosophila melanogaster. Genetics. 1999 Feb;151(2):725–737. doi: 10.1093/genetics/151.2.725. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. Tomlinson A., Bowtell D. D., Hafen E., Rubin G. M. Localization of the sevenless protein, a putative receptor for positional information, in the eye imaginal disc of Drosophila. Cell. 1987 Oct 9;51(1):143–150. doi: 10.1016/0092-8674(87)90019-5. [DOI] [PubMed] [Google Scholar]
  45. Treisman J. E., Ito N., Rubin G. M. misshapen encodes a protein kinase involved in cell shape control in Drosophila. Gene. 1997 Feb 20;186(1):119–125. doi: 10.1016/s0378-1119(96)00694-4. [DOI] [PubMed] [Google Scholar]
  46. Vossler M. R., Yao H., York R. D., Pan M. G., Rim C. S., Stork P. J. cAMP activates MAP kinase and Elk-1 through a B-Raf- and Rap1-dependent pathway. Cell. 1997 Apr 4;89(1):73–82. doi: 10.1016/s0092-8674(00)80184-1. [DOI] [PubMed] [Google Scholar]
  47. Yu W., Fantl W. J., Harrowe G., Williams L. T. Regulation of the MAP kinase pathway by mammalian Ksr through direct interaction with MEK and ERK. Curr Biol. 1998 Jan 1;8(1):56–64. doi: 10.1016/s0960-9822(98)70020-x. [DOI] [PubMed] [Google Scholar]
  48. 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]

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