Cloning and characterization of a protein kinase C gene from the spruce budworm, Choristoneura fumiferana.
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.
Quan
Guo-Xing
GX
Great Lakes Forestry Centre, Canadian Forest Service, 1219 Queen Street East, Sault Ste. Marie, Ontario, Canada P6A 2E5. gquan@nrcan.gc.ca
Doucet
Daniel
D
Ladd
Tim
T
Krell
Peter J
PJ
Arif
Basil M
BM
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002478:Cells, Cultured; D003001:Cloning, Molecular; D004789:Enzyme Activation; D015870:Gene Expression; D049452:Green Fluorescent Proteins / Q000032:analysis; D019476:Insect Proteins / Q000032:analysis / Q000737:chemistry / Q000235:genetics*; D008969:Molecular Sequence Data; D009036:Moths / Q000187:drug effects / Q000201:enzymology* / Q000235:genetics; D011493:Protein Kinase C / Q000032:analysis / Q000737:chemistry / Q000235:genetics*; D034622:RNA Interference; D012333:RNA, Messenger / Q000378:metabolism; D017422:Sequence Analysis, DNA; D020539:Sequence Analysis, Protein; D015398:Signal Transduction / Q000235:genetics; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology
D019476:Insect Proteins; D012333:RNA, Messenger; D049452:Green Fluorescent Proteins; D011493:Protein Kinase C; D013755:Tetradecanoylphorbol Acetate