Lipopolysaccharide-activated kinase, an essential component for the induction of the antimicrobial peptide genes in Drosophila melanogaster cells

Yong Sik Kim, Sung Jun Han, Ji Hwan Ryu, Kun Ho Choi, Young Suk Hong, Yong-Hoon Chung, Sylvie Perrot, Anna Raibaud, Paul T. Brey, Won Jae Lee

Research output: Contribution to journalArticle

33 Citations (Scopus)

Abstract

Eukaryotic organisms use a similar Rel/NF-κB signaling cascade for the induction of innate immune genes. In Drosophila, lipopolysaccharide (LPS) signal-induced activation of the Rel/NF-κB family transcription factors is an essential step in the transcriptional activation of inducible antimicrobial peptide genes. However, the mechanism by which the LPS-induced signaling pathway proceeds remains largely unknown. Here we have cloned a novel Drosophila LPS-activated kinase (DLAK) that is structurally related to mammalian IκB kinases. DLAK is expressed and transiently activated in LPS- responsive Drosophila cells following LPS stimulation. Furthermore, DLAK can interact with Cactus, a Drosophila IκB and phosphorylate recombinant Cactus, in vitro. Overexpression of dominant-negative mutant DLAK (DLAK(K50A)) blocks LPS-induced Cactus degradation. DLAK-bound Cactus can be degraded in a LPS signal-dependent fashion, whereas the DLAK(K50A) mutant-bound Cactus is completely resistant to degradation in the presence of LPS. The DLAK(K50A) mutant also inhibits nuclear κB binding activity and κB-dependent diptericin reporter gene activity in a dose-dependent manner, but the κB- dependent diptericin reporter gene activity can be rescued by overexpression of wild type DLAK. Moreover, mRNA analysis of various κB-dependent antimicrobial peptide genes shows that LPS inducibility of these genes is greatly impaired in cells overexpressing DLAK(K50A). These results establish that DLAK is a novel LPS-activated kinase, which is an essential signaling component for the induction of antimicrobial peptide genes following LPS treatment in Drosophila cells.

Original languageEnglish
Pages (from-to)2071-2079
Number of pages9
JournalJournal of Biological Chemistry
Volume275
Issue number3
DOIs
StatePublished - 2000 Jan 21

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Drosophila melanogaster
Lipopolysaccharides
Phosphotransferases
Genes
Cactaceae
Peptides
Drosophila
Reporter Genes
Drosophila ird5 protein
Chemical activation
Degradation
Transcriptional Activation
Transcription Factors
Messenger RNA

Cite this

Kim, Yong Sik ; Han, Sung Jun ; Ryu, Ji Hwan ; Choi, Kun Ho ; Hong, Young Suk ; Chung, Yong-Hoon ; Perrot, Sylvie ; Raibaud, Anna ; Brey, Paul T. ; Lee, Won Jae. / Lipopolysaccharide-activated kinase, an essential component for the induction of the antimicrobial peptide genes in Drosophila melanogaster cells. In: Journal of Biological Chemistry. 2000 ; Vol. 275, No. 3. pp. 2071-2079.
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abstract = "Eukaryotic organisms use a similar Rel/NF-κB signaling cascade for the induction of innate immune genes. In Drosophila, lipopolysaccharide (LPS) signal-induced activation of the Rel/NF-κB family transcription factors is an essential step in the transcriptional activation of inducible antimicrobial peptide genes. However, the mechanism by which the LPS-induced signaling pathway proceeds remains largely unknown. Here we have cloned a novel Drosophila LPS-activated kinase (DLAK) that is structurally related to mammalian IκB kinases. DLAK is expressed and transiently activated in LPS- responsive Drosophila cells following LPS stimulation. Furthermore, DLAK can interact with Cactus, a Drosophila IκB and phosphorylate recombinant Cactus, in vitro. Overexpression of dominant-negative mutant DLAK (DLAK(K50A)) blocks LPS-induced Cactus degradation. DLAK-bound Cactus can be degraded in a LPS signal-dependent fashion, whereas the DLAK(K50A) mutant-bound Cactus is completely resistant to degradation in the presence of LPS. The DLAK(K50A) mutant also inhibits nuclear κB binding activity and κB-dependent diptericin reporter gene activity in a dose-dependent manner, but the κB- dependent diptericin reporter gene activity can be rescued by overexpression of wild type DLAK. Moreover, mRNA analysis of various κB-dependent antimicrobial peptide genes shows that LPS inducibility of these genes is greatly impaired in cells overexpressing DLAK(K50A). These results establish that DLAK is a novel LPS-activated kinase, which is an essential signaling component for the induction of antimicrobial peptide genes following LPS treatment in Drosophila cells.",
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Lipopolysaccharide-activated kinase, an essential component for the induction of the antimicrobial peptide genes in Drosophila melanogaster cells. / Kim, Yong Sik; Han, Sung Jun; Ryu, Ji Hwan; Choi, Kun Ho; Hong, Young Suk; Chung, Yong-Hoon; Perrot, Sylvie; Raibaud, Anna; Brey, Paul T.; Lee, Won Jae.

In: Journal of Biological Chemistry, Vol. 275, No. 3, 21.01.2000, p. 2071-2079.

Research output: Contribution to journalArticle

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AU - Kim, Yong Sik

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AU - Ryu, Ji Hwan

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AU - Hong, Young Suk

AU - Chung, Yong-Hoon

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AU - Raibaud, Anna

AU - Brey, Paul T.

AU - Lee, Won Jae

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