Abstract
Eight alleles of Dsor1 encoding a Drosophila homologue of mitogen-activated protein (MAP) kinase kinase were obtained as dominant suppressors of the MAP kinase kinase kinase D-raf. These Dsor1 alleles themselves showed no obvious phenotypic consequences nor any effect on the viability of the flies, although they were highly sensitive to upstream signals and strongly interacted with gain-of-function mutations of upstream factors. They suppressed mutations for receptor tyrosine kinases (RTKs); torso (tor), sevenless (sev) and to a lesser extent Drosophila EGF receptor (DER). Furthermore, the Dsor1 alleles showed no significant interaction with gain-of-function mutations of DER. The observed difference in activity of the Dsor1 alleles among the RTK pathways suggests Dsor1 is one of the components of the pathway that regulates signal specificity. Expression of Dsor1 in budding yeast demonstrated that Dsor1 can activate yeast MAP kinase homologues if a proper activator of Dsor1 is coexpressed. Nucleotide sequencing of the Dsor1 mutant genes revealed that most of the mutations are associated with amino acid changes at highly conserved residues in the kinase domain. The results suggest that they function as suppressors due to increased reactivity to upstream factors.
Full Text
The Full Text of this article is available as a PDF (5.7 MB).
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- 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., 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]
- Casanova J., Llimargas M., Greenwood S., Struhl G. An oncogenic form of human raf can specify terminal body pattern in Drosophila. Mech Dev. 1994 Oct;48(1):59–64. doi: 10.1016/0925-4773(94)90006-x. [DOI] [PubMed] [Google Scholar]
- Catling A. D., Schaeffer H. J., Reuter C. W., Reddy G. R., Weber M. J. A proline-rich sequence unique to MEK1 and MEK2 is required for raf binding and regulates MEK function. Mol Cell Biol. 1995 Oct;15(10):5214–5225. doi: 10.1128/mcb.15.10.5214. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Choi K. Y., Satterberg B., Lyons D. M., Elion E. A. Ste5 tethers multiple protein kinases in the MAP kinase cascade required for mating in S. cerevisiae. Cell. 1994 Aug 12;78(3):499–512. doi: 10.1016/0092-8674(94)90427-8. [DOI] [PubMed] [Google Scholar]
- Chuang E., Barnard D., Hettich L., Zhang X. F., Avruch J., Marshall M. S. Critical binding and regulatory interactions between Ras and Raf occur through a small, stable N-terminal domain of Raf and specific Ras effector residues. Mol Cell Biol. 1994 Aug;14(8):5318–5325. doi: 10.1128/mcb.14.8.5318. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Clifford R., Schüpbach T. Molecular analysis of the Drosophila EGF receptor homolog reveals that several genetically defined classes of alleles cluster in subdomains of the receptor protein. Genetics. 1994 Jun;137(2):531–550. doi: 10.1093/genetics/137.2.531. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Crews C. M., Alessandrini A., Erikson R. L. The primary structure of MEK, a protein kinase that phosphorylates the ERK gene product. Science. 1992 Oct 16;258(5081):478–480. doi: 10.1126/science.1411546. [DOI] [PubMed] [Google Scholar]
- Crews C. M., Erikson R. L. Purification of a murine protein-tyrosine/threonine kinase that phosphorylates and activates the Erk-1 gene product: relationship to the fission yeast byr1 gene product. Proc Natl Acad Sci U S A. 1992 Sep 1;89(17):8205–8209. doi: 10.1073/pnas.89.17.8205. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Davis R. J. MAPKs: new JNK expands the group. Trends Biochem Sci. 1994 Nov;19(11):470–473. doi: 10.1016/0968-0004(94)90132-5. [DOI] [PubMed] [Google Scholar]
- 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]
- Dickson B., Sprenger F., Morrison D., Hafen E. Raf functions downstream of Ras1 in the Sevenless signal transduction pathway. Nature. 1992 Dec 10;360(6404):600–603. doi: 10.1038/360600a0. [DOI] [PubMed] [Google Scholar]
- Fabian J. R., Vojtek A. B., Cooper J. A., Morrison D. K. A single amino acid change in Raf-1 inhibits Ras binding and alters Raf-1 function. Proc Natl Acad Sci U S A. 1994 Jun 21;91(13):5982–5986. doi: 10.1073/pnas.91.13.5982. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 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]
- Hanks S. K., Quinn A. M., Hunter T. The protein kinase family: conserved features and deduced phylogeny of the catalytic domains. Science. 1988 Jul 1;241(4861):42–52. doi: 10.1126/science.3291115. [DOI] [PubMed] [Google Scholar]
- Hata M., Inoue Y. H., Yoo M. A., Nishida Y. Multiple functions of raf proto-oncogene during development from analysis of a temperature-sensitive mutation of Drosophila. Int J Dev Biol. 1994 Jun;38(2):329–335. [PubMed] [Google Scholar]
- Hsu J. C., Perrimon N. A temperature-sensitive MEK mutation demonstrates the conservation of the signaling pathways activated by receptor tyrosine kinases. Genes Dev. 1994 Sep 15;8(18):2176–2187. doi: 10.1101/gad.8.18.2176. [DOI] [PubMed] [Google Scholar]
- Huang W., Erikson R. L. Constitutive activation of Mek1 by mutation of serine phosphorylation sites. Proc Natl Acad Sci U S A. 1994 Sep 13;91(19):8960–8963. doi: 10.1073/pnas.91.19.8960. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Irie K., Gotoh Y., Yashar B. M., Errede B., Nishida E., Matsumoto K. Stimulatory effects of yeast and mammalian 14-3-3 proteins on the Raf protein kinase. Science. 1994 Sep 16;265(5179):1716–1719. doi: 10.1126/science.8085159. [DOI] [PubMed] [Google Scholar]
- Irie K., Takase M., Lee K. S., Levin D. E., Araki H., Matsumoto K., Oshima Y. MKK1 and MKK2, which encode Saccharomyces cerevisiae mitogen-activated protein kinase-kinase homologs, function in the pathway mediated by protein kinase C. Mol Cell Biol. 1993 May;13(5):3076–3083. doi: 10.1128/mcb.13.5.3076. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Johnson L. N., Noble M. E., Owen D. J. Active and inactive protein kinases: structural basis for regulation. Cell. 1996 Apr 19;85(2):149–158. doi: 10.1016/s0092-8674(00)81092-2. [DOI] [PubMed] [Google Scholar]
- Koga M., Ohshima Y. Drosophila MAP kinase kinase suppresses the vulvaless phenotype of lin-3, let-23 and lin-45 mutations in Caenorhabditis elegans. Mech Dev. 1995 Sep;53(1):15–22. doi: 10.1016/0925-4773(95)00417-3. [DOI] [PubMed] [Google Scholar]
- Lu X., Melnick M. B., Hsu J. C., Perrimon N. Genetic and molecular analyses of mutations involved in Drosophila raf signal transduction. EMBO J. 1994 Jun 1;13(11):2592–2599. doi: 10.1002/j.1460-2075.1994.tb06549.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Maeda T., Takekawa M., Saito H. Activation of yeast PBS2 MAPKK by MAPKKKs or by binding of an SH3-containing osmosensor. Science. 1995 Jul 28;269(5223):554–558. doi: 10.1126/science.7624781. [DOI] [PubMed] [Google Scholar]
- Neiman A. M. Conservation and reiteration of a kinase cascade. Trends Genet. 1993 Nov;9(11):390–394. doi: 10.1016/0168-9525(93)90139-9. [DOI] [PubMed] [Google Scholar]
- Nishida E., Gotoh Y. The MAP kinase cascade is essential for diverse signal transduction pathways. Trends Biochem Sci. 1993 Apr;18(4):128–131. doi: 10.1016/0968-0004(93)90019-j. [DOI] [PubMed] [Google Scholar]
- 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]
- 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]
- Price J. V., Clifford R. J., Schüpbach T. The maternal ventralizing locus torpedo is allelic to faint little ball, an embryonic lethal, and encodes the Drosophila EGF receptor homolog. Cell. 1989 Mar 24;56(6):1085–1092. doi: 10.1016/0092-8674(89)90641-7. [DOI] [PubMed] [Google Scholar]
- Rogge R. D., Karlovich C. A., Banerjee U. Genetic dissection of a neurodevelopmental pathway: Son of sevenless functions downstream of the sevenless and EGF receptor tyrosine kinases. Cell. 1991 Jan 11;64(1):39–48. doi: 10.1016/0092-8674(91)90207-f. [DOI] [PubMed] [Google Scholar]
- Rossomando A., Wu J., Weber M. J., Sturgill T. W. The phorbol ester-dependent activator of the mitogen-activated protein kinase p42mapk is a kinase with specificity for the threonine and tyrosine regulatory sites. Proc Natl Acad Sci U S A. 1992 Jun 15;89(12):5221–5225. doi: 10.1073/pnas.89.12.5221. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Scott A., Haystead C. M., Haystead T. A. Purification of a 12,020-dalton protein that enhances the activation of mitogen-activated protein (MAP) kinase by MAP kinase kinase. J Biol Chem. 1995 Oct 13;270(41):24540–24547. doi: 10.1074/jbc.270.41.24540. [DOI] [PubMed] [Google Scholar]
- Sherman F. Getting started with yeast. Methods Enzymol. 1991;194:3–21. doi: 10.1016/0076-6879(91)94004-v. [DOI] [PubMed] [Google Scholar]
- Stevenson B. J., Rhodes N., Errede B., Sprague G. F., Jr Constitutive mutants of the protein kinase STE11 activate the yeast pheromone response pathway in the absence of the G protein. Genes Dev. 1992 Jul;6(7):1293–1304. doi: 10.1101/gad.6.7.1293. [DOI] [PubMed] [Google Scholar]
- Teague M. A., Chaleff D. T., Errede B. Nucleotide sequence of the yeast regulatory gene STE7 predicts a protein homologous to protein kinases. Proc Natl Acad Sci U S A. 1986 Oct;83(19):7371–7375. doi: 10.1073/pnas.83.19.7371. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 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]
- Vojtek A. B., Hollenberg S. M., Cooper J. A. Mammalian Ras interacts directly with the serine/threonine kinase Raf. Cell. 1993 Jul 16;74(1):205–214. doi: 10.1016/0092-8674(93)90307-c. [DOI] [PubMed] [Google Scholar]
- Whiteway M., Hougan L., Dignard D., Thomas D. Y., Bell L., Saari G. C., Grant F. J., O'Hara P., MacKay V. L. The STE4 and STE18 genes of yeast encode potential beta and gamma subunits of the mating factor receptor-coupled G protein. Cell. 1989 Feb 10;56(3):467–477. doi: 10.1016/0092-8674(89)90249-3. [DOI] [PubMed] [Google Scholar]
- Yan M., Templeton D. J. Identification of 2 serine residues of MEK-1 that are differentially phosphorylated during activation by raf and MEK kinase. J Biol Chem. 1994 Jul 22;269(29):19067–19073. [PubMed] [Google Scholar]
