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

Insect Biochem Mol Biol. 2008 Nov;38(11):984-92. doi: 10.1016/j.ibmb.2008.08.003. Epub 2008 Aug 26.

Abstract

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Base Sequence
  • Cells, Cultured
  • Cloning, Molecular
  • Enzyme Activation
  • Gene Expression
  • Green Fluorescent Proteins / analysis
  • Insect Proteins / analysis
  • Insect Proteins / chemistry
  • Insect Proteins / genetics*
  • Molecular Sequence Data
  • Moths / drug effects
  • Moths / enzymology*
  • Moths / genetics
  • Protein Kinase C / analysis
  • Protein Kinase C / chemistry
  • Protein Kinase C / genetics*
  • RNA Interference
  • RNA, Messenger / metabolism
  • Sequence Analysis, DNA
  • Sequence Analysis, Protein
  • Signal Transduction / genetics
  • Tetradecanoylphorbol Acetate / pharmacology

Substances

  • Insect Proteins
  • RNA, Messenger
  • Green Fluorescent Proteins
  • Protein Kinase C
  • Tetradecanoylphorbol Acetate