Molecular Cloning and Characterization of aDrosophila p38 Mitogen-activated Protein Kinase*
- From the ‡Laboratory of Immunology, Medical Research Center, College of Medicine, Yonsei University, CPO Box 8044, Seoul, South Korea, the §Department of Biochemistry and Molecular Biology, College of Medicine, Yonsei University, CPO Box 8044, Seoul, South Korea, and the ¶Laboratoire de Biochimie et Biologie Moléculaire des Insectes, Institut Pasteur, 75724, Paris Cedex 15, Paris, France
Abstract
A mitogen-activated protein kinase (MAPK) has been cloned and sequenced from a Drosophila neoplasmicl(2)mbn cell line. The cDNA sequence analysis showed that this Drosophila kinase is a homologue of mammalian p38 MAPK and the yeast HOG1 gene and thus was referred to as Dp38. A distinguishing feature of all MAPKs is the conserved sequence TGY in the activation domain. Dp38 was rapidly tyrosine 186-phosphorylated in response to osmotic stress, heat shock, serum starvation, and H2O2 in Drosophila l(2)mbn and Schneider cell lines. However, unlike mammalian p38 MAPK, the addition of lipopolysaccharide (LPS) did not significantly affect the phosphorylation of Dp38 in the LPS-responsivel(2)mbn cell line. Following osmotic stress, tyrosine 186-phosphorylated forms of Dp38 MAPK were detected exclusively in nuclear regions of Schneider cells. Yeast complementation studies demonstrated that the Saccharomyces cerevisiae HOG1 mutant strain JBY10 (hog1-Δ1) was functionally complemented by Dp38 cDNA in hyperosmolar medium. These findings demonstrate that similar osmotic stress-responsive signal transduction pathways are conserved in yeast, Drosophila, and mammalian cells, whereas LPS signal transduction pathways appear to be different.
Footnotes
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↵* This work has been supported by Genetic Engineering Research Grants GE 96–128 and GE 97–114 from the Korea Ministry of Education (to W.-J. L.) and partly by the Yonsei University College of Medicine Faculty Grant (1996), the Yonsei University Research Fund (1996), a grant from the Hallym Academy of Science (to K.-Y. C.), and grants from Institut Pasteur and the Pasteur-Weizmann Joint Research Program.The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked “advertisement” in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.
The nucleotide sequence(s) reported in this paper has been submitted to the GenBank™/EMBL Data Bank with accession number(s) U86867.
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↵‖ To whom correspondence should be addressed: Laboratory of Immunology, Medical Research Center, College of Medicine, Yonsei University, CPO Box 8044, Seoul, South Korea. Tel.: 82-23618339; Fax: 82-23628647; E-mail: wjlee1{at}yumc.yonsei.ac.kr.
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↵1 The abbreviations used are: MAPK, mitogen-activated protein kinase; HOG1, high osmolarity glycerol protein kinase in S. cerevisiae; LPS, lipopolysaccharide; RK, reactivating kinase; JNK, c-Jun NH2-terminal kinase; ERK, extracellular signal-regulated kinase; MKK, MAPK kinase; PCR, polymerase chain reaction; PAGE, polyacrylamide gel electrophoresis; bp, base pair(s); SC, synthetic complete.
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- Received September 11, 1997.
- Revision received October 26, 1997.
- The American Society for Biochemistry and Molecular Biology, Inc.
