Abstract
The Drosophila nonreceptor protein tyrosine phosphatase, Corkscrew (Csw), functions positively in multiple receptor tyrosine kinase (RTK) pathways, including signaling by the epidermal growth factor receptor (EGFR). Detailed phenotypic analyses of csw mutations have revealed that Csw activity is required in many of the same developmental processes that require EGFR function. However, it is still unclear where in the signaling hierarchy Csw functions relative to other proteins whose activities are also required downstream of the receptor. To address this issue, genetic interaction experiments were performed to place csw gene activity relative to the EGFR, spitz (spi), rhomboid (rho), daughter of sevenless (DOS), kinase-suppressor of ras (ksr), ras1, D-raf, pointed (pnt), and moleskin. We followed the EGFR-dependent formation of VA2 muscle precursor cells as a sensitive assay for these genetic interaction studies. First, we established that Csw has a positive function during mesoderm development. Second, we found that tissue-specific expression of a gain-of-function csw construct rescues loss-of-function mutations in other positive signaling genes upstream of rolled (rl)/MAPK in the EGFR pathway. Third, we were able to infer levels of EGFR signaling in various mutant backgrounds during myogenesis. This work extends previous studies of Csw during Torso and Sevenless RTK signaling to include an in-depth analysis of the role of Csw in the EGFR signaling pathway.
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- Allard J. D., Chang H. C., Herbst R., McNeill H., Simon M. A. The SH2-containing tyrosine phosphatase corkscrew is required during signaling by sevenless, Ras1 and Raf. Development. 1996 Apr;122(4):1137–1146. doi: 10.1242/dev.122.4.1137. [DOI] [PubMed] [Google Scholar]
- 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]
- Bausenwein B. S., Schmidt M., Mielke B., Raabe T. In vivo functional analysis of the daughter of sevenless protein in receptor tyrosine kinase signaling. Mech Dev. 2000 Feb;90(2):205–215. doi: 10.1016/s0925-4773(99)00252-x. [DOI] [PubMed] [Google Scholar]
- Brand A. H., Perrimon N. Raf acts downstream of the EGF receptor to determine dorsoventral polarity during Drosophila oogenesis. Genes Dev. 1994 Mar 1;8(5):629–639. doi: 10.1101/gad.8.5.629. [DOI] [PubMed] [Google Scholar]
- Brunner D., Dücker K., Oellers N., Hafen E., Scholz H., Klämbt C. The ETS domain protein pointed-P2 is a target of MAP kinase in the sevenless signal transduction pathway. Nature. 1994 Aug 4;370(6488):386–389. doi: 10.1038/370386a0. [DOI] [PubMed] [Google Scholar]
- Brunner D., Oellers N., Szabad J., Biggs W. H., 3rd, Zipursky S. L., Hafen E. A gain-of-function mutation in Drosophila MAP kinase activates multiple receptor tyrosine kinase signaling pathways. Cell. 1994 Mar 11;76(5):875–888. doi: 10.1016/0092-8674(94)90362-x. [DOI] [PubMed] [Google Scholar]
- Buff E., Carmena A., Gisselbrecht S., Jiménez F., Michelson A. M. Signalling by the Drosophila epidermal growth factor receptor is required for the specification and diversification of embryonic muscle progenitors. Development. 1998 Jun;125(11):2075–2086. doi: 10.1242/dev.125.11.2075. [DOI] [PubMed] [Google Scholar]
- Carmena A., Gisselbrecht S., Harrison J., Jiménez F., Michelson A. M. Combinatorial signaling codes for the progressive determination of cell fates in the Drosophila embryonic mesoderm. Genes Dev. 1998 Dec 15;12(24):3910–3922. doi: 10.1101/gad.12.24.3910. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Casci T., Freeman M. Control of EGF receptor signalling: lessons from fruitflies. Cancer Metastasis Rev. 1999;18(2):181–201. doi: 10.1023/a:1006313122373. [DOI] [PubMed] [Google Scholar]
- 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]
- Cleghon V., Feldmann P., Ghiglione C., Copeland T. D., Perrimon N., Hughes D. A., Morrison D. K. Opposing actions of CSW and RasGAP modulate the strength of Torso RTK signaling in the Drosophila terminal pathway. Mol Cell. 1998 Dec;2(6):719–727. doi: 10.1016/s1097-2765(00)80287-7. [DOI] [PubMed] [Google Scholar]
- Cleghon V., Gayko U., Copeland T. D., Perkins L. A., Perrimon N., Morrison D. K. Drosophila terminal structure development is regulated by the compensatory activities of positive and negative phosphotyrosine signaling sites on the Torso RTK. Genes Dev. 1996 Mar 1;10(5):566–577. doi: 10.1101/gad.10.5.566. [DOI] [PubMed] [Google Scholar]
- 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]
- Freeman M., Kimmel B. E., Rubin G. M. Identifying targets of the rough homeobox gene of Drosophila: evidence that rhomboid functions in eye development. Development. 1992 Oct;116(2):335–346. doi: 10.1242/dev.116.2.335. [DOI] [PubMed] [Google Scholar]
- Garrington T. P., Johnson G. L. Organization and regulation of mitogen-activated protein kinase signaling pathways. Curr Opin Cell Biol. 1999 Apr;11(2):211–218. doi: 10.1016/s0955-0674(99)80028-3. [DOI] [PubMed] [Google Scholar]
- Gaul U., Seifert E., Schuh R., Jäckle H. Analysis of Krüppel protein distribution during early Drosophila development reveals posttranscriptional regulation. Cell. 1987 Aug 14;50(4):639–647. doi: 10.1016/0092-8674(87)90037-7. [DOI] [PubMed] [Google Scholar]
- Greig S., Akam M. Homeotic genes autonomously specify one aspect of pattern in the Drosophila mesoderm. Nature. 1993 Apr 15;362(6421):630–632. doi: 10.1038/362630a0. [DOI] [PubMed] [Google Scholar]
- Halfon M. S., Carmena A., Gisselbrecht S., Sackerson C. M., Jiménez F., Baylies M. K., Michelson A. M. Ras pathway specificity is determined by the integration of multiple signal-activated and tissue-restricted transcription factors. Cell. 2000 Sep 29;103(1):63–74. doi: 10.1016/s0092-8674(00)00105-7. [DOI] [PubMed] [Google Scholar]
- Herbst R., Carroll P. M., Allard J. D., Schilling J., Raabe T., Simon M. A. Daughter of sevenless is a substrate of the phosphotyrosine phosphatase Corkscrew and functions during sevenless signaling. Cell. 1996 Jun 14;85(6):899–909. doi: 10.1016/s0092-8674(00)81273-8. [DOI] [PubMed] [Google Scholar]
- Herbst R., Zhang X., Qin J., Simon M. A. Recruitment of the protein tyrosine phosphatase CSW by DOS is an essential step during signaling by the sevenless receptor tyrosine kinase. EMBO J. 1999 Dec 15;18(24):6950–6961. doi: 10.1093/emboj/18.24.6950. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hof P., Pluskey S., Dhe-Paganon S., Eck M. J., Shoelson S. E. Crystal structure of the tyrosine phosphatase SHP-2. Cell. 1998 Feb 20;92(4):441–450. doi: 10.1016/s0092-8674(00)80938-1. [DOI] [PubMed] [Google Scholar]
- Holgado-Madruga M., Emlet D. R., Moscatello D. K., Godwin A. K., Wong A. J. A Grb2-associated docking protein in EGF- and insulin-receptor signalling. Nature. 1996 Feb 8;379(6565):560–564. doi: 10.1038/379560a0. [DOI] [PubMed] [Google Scholar]
- Hou X. S., Chou T. B., Melnick M. B., Perrimon N. The torso receptor tyrosine kinase can activate Raf in a Ras-independent pathway. Cell. 1995 Apr 7;81(1):63–71. doi: 10.1016/0092-8674(95)90371-2. [DOI] [PubMed] [Google Scholar]
- Klämbt C. The Drosophila gene pointed encodes two ETS-like proteins which are involved in the development of the midline glial cells. Development. 1993 Jan;117(1):163–176. doi: 10.1242/dev.117.1.163. [DOI] [PubMed] [Google Scholar]
- Lee T., Feig L., Montell D. J. Two distinct roles for Ras in a developmentally regulated cell migration. Development. 1996 Feb;122(2):409–418. doi: 10.1242/dev.122.2.409. [DOI] [PubMed] [Google Scholar]
- Lehr S., Kotzka J., Herkner A., Klein E., Siethoff C., Knebel B., Noelle V., Brüning J. C., Klein H. W., Meyer H. E. Identification of tyrosine phosphorylation sites in human Gab-1 protein by EGF receptor kinase in vitro. Biochemistry. 1999 Jan 5;38(1):151–159. doi: 10.1021/bi9818265. [DOI] [PubMed] [Google Scholar]
- Li W., Skoulakis E. M., Davis R. L., Perrimon N. The Drosophila 14-3-3 protein Leonardo enhances Torso signaling through D-Raf in a Ras 1-dependent manner. Development. 1997 Oct;124(20):4163–4171. doi: 10.1242/dev.124.20.4163. [DOI] [PubMed] [Google Scholar]
- Lorenzen J. A., Baker S. E., Denhez F., Melnick M. B., Brower D. L., Perkins L. A. Nuclear import of activated D-ERK by DIM-7, an importin family member encoded by the gene moleskin. Development. 2001 Apr;128(8):1403–1414. doi: 10.1242/dev.128.8.1403. [DOI] [PubMed] [Google Scholar]
- Michelson A. M., Gisselbrecht S., Zhou Y., Baek K. H., Buff E. M. Dual functions of the heartless fibroblast growth factor receptor in development of the Drosophila embryonic mesoderm. Dev Genet. 1998;22(3):212–229. doi: 10.1002/(SICI)1520-6408(1998)22:3<212::AID-DVG4>3.0.CO;2-9. [DOI] [PubMed] [Google Scholar]
- 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]
- Perkins L. A., Johnson M. R., Melnick M. B., Perrimon N. The nonreceptor protein tyrosine phosphatase corkscrew functions in multiple receptor tyrosine kinase pathways in Drosophila. Dev Biol. 1996 Nov 25;180(1):63–81. doi: 10.1006/dbio.1996.0285. [DOI] [PubMed] [Google Scholar]
- Perkins L. A., Larsen I., Perrimon N. corkscrew encodes a putative protein tyrosine phosphatase that functions to transduce the terminal signal from the receptor tyrosine kinase torso. Cell. 1992 Jul 24;70(2):225–236. doi: 10.1016/0092-8674(92)90098-w. [DOI] [PubMed] [Google Scholar]
- Perrimon N., Perkins L. A. There must be 50 ways to rule the signal: the case of the Drosophila EGF receptor. Cell. 1997 Apr 4;89(1):13–16. doi: 10.1016/s0092-8674(00)80177-4. [DOI] [PubMed] [Google Scholar]
- Queenan A. M., Ghabrial A., Schüpbach T. Ectopic activation of torpedo/Egfr, a Drosophila receptor tyrosine kinase, dorsalizes both the eggshell and the embryo. Development. 1997 Oct;124(19):3871–3880. doi: 10.1242/dev.124.19.3871. [DOI] [PubMed] [Google Scholar]
- Raabe T., Riesgo-Escovar J., Liu X., Bausenwein B. S., Deak P., Maröy P., Hafen E. DOS, a novel pleckstrin homology domain-containing protein required for signal transduction between sevenless and Ras1 in Drosophila. Cell. 1996 Jun 14;85(6):911–920. doi: 10.1016/s0092-8674(00)81274-x. [DOI] [PubMed] [Google Scholar]
- Raabe T. The sevenless signaling pathway: variations of a common theme. Biochim Biophys Acta. 2000 Apr 17;1496(2-3):151–163. doi: 10.1016/s0167-4889(00)00020-3. [DOI] [PubMed] [Google Scholar]
- Rodrigues G. A., Falasca M., Zhang Z., Ong S. H., Schlessinger J. A novel positive feedback loop mediated by the docking protein Gab1 and phosphatidylinositol 3-kinase in epidermal growth factor receptor signaling. Mol Cell Biol. 2000 Feb;20(4):1448–1459. doi: 10.1128/mcb.20.4.1448-1459.2000. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Rommel C., Hafen E. Ras--a versatile cellular switch. Curr Opin Genet Dev. 1998 Aug;8(4):412–418. doi: 10.1016/s0959-437x(98)80111-1. [DOI] [PubMed] [Google Scholar]
- Ruohola-Baker H., Grell E., Chou T. B., Baker D., Jan L. Y., Jan Y. N. Spatially localized rhomboid is required for establishment of the dorsal-ventral axis in Drosophila oogenesis. Cell. 1993 Jun 4;73(5):953–965. doi: 10.1016/0092-8674(93)90273-s. [DOI] [PubMed] [Google Scholar]
- Rutledge B. J., Zhang K., Bier E., Jan Y. N., Perrimon N. The Drosophila spitz gene encodes a putative EGF-like growth factor involved in dorsal-ventral axis formation and neurogenesis. Genes Dev. 1992 Aug;6(8):1503–1517. doi: 10.1101/gad.6.8.1503. [DOI] [PubMed] [Google Scholar]
- Schlessinger J. Cell signaling by receptor tyrosine kinases. Cell. 2000 Oct 13;103(2):211–225. doi: 10.1016/s0092-8674(00)00114-8. [DOI] [PubMed] [Google Scholar]
- Schweitzer R., Shilo B. Z. A thousand and one roles for the Drosophila EGF receptor. Trends Genet. 1997 May;13(5):191–196. doi: 10.1016/s0168-9525(97)01091-3. [DOI] [PubMed] [Google Scholar]
- Sturtevant M. A., Roark M., Bier E. The Drosophila rhomboid gene mediates the localized formation of wing veins and interacts genetically with components of the EGF-R signaling pathway. Genes Dev. 1993 Jun;7(6):961–973. doi: 10.1101/gad.7.6.961. [DOI] [PubMed] [Google Scholar]
- Suzanne M., Irie K., Glise B., Agnès F., Mori E., Matsumoto K., Noselli S. The Drosophila p38 MAPK pathway is required during oogenesis for egg asymmetric development. Genes Dev. 1999 Jun 1;13(11):1464–1474. doi: 10.1101/gad.13.11.1464. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 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]
- 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]
- Yarnitzky T., Min L., Volk T. The Drosophila neuregulin homolog Vein mediates inductive interactions between myotubes and their epidermal attachment cells. Genes Dev. 1997 Oct 15;11(20):2691–2700. doi: 10.1101/gad.11.20.2691. [DOI] [PMC free article] [PubMed] [Google Scholar]