Cloning and characterization of a protein kinase C gene from the spruce budworm, Choristoneura fumiferana

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• 作者：Guo-Xing Quan ; Daniel Doucet ; Tim Ladd ; Peter J. Krell ; Basil M. Arif
• 关键词：Phosphorylation ; Signal transduction ; Insect molting
• 刊名：Insect Biochemistry and Molecular Biology
• 出版年：2008
• 出版时间：November 2008
• 年：2008
• 卷：38
• 期：11
• 页码：984-992
• 全文大小：1465 K

文摘

Recent studies have implicated protein kinase C (PKC) in the control of 20-hydroxyecdysone (20E)-dependent gene expression during molting and metamorphosis in insects. To further understand the role of this kinase in 20E signal transduction, we cloned a homolog of mammalian PKC by RT-PCR and 5′/3′-RACE from adult of the moth Choristoneura fumiferana. The full-length cDNA of the C. fumiferana PKC (CfPKC1) is 2.3 kb with an open reading frame encoding a protein of 669 amino acids. The deduced amino acid sequence contains all the characteristic features of the classical protein kinase C subfamily. Northern and Western blot analysis showed that CfPKC1 was distributed ubiquitously in various tissues and at different developmental stages. Activation of CfPKC1 with the PKC activator phorbol 12-myristate 13-acetate (PMA) resulted in a rapid redistribution of the protein from the cytosol to the plasma membrane. Knock-down of the CfPKC1 gene by double-stranded RNA interference or treatment of the CF-203 cells with PKC-specific inhibitors reduces the expression of the 20E-responsive genes CHR3 and E75. This data suggests that CfPKC1 is involved in the 20E-response gene expression in C. fumiferana.