Receptor of activated C kinase 1 (RACK1) is necessary for the 20-hydroxyecdysone-induced expression of the transcription factor CHR3 in the spruce budworm Choristoneura fumiferana.
To initiate moulting and metamorphosis, 20-hydroxyecdysone (20E) binds to its nuclear receptors and the ligand-receptor complex then mediates changes in gene expression. Phosphorylation of the receptors is required for their function. The intracellular signal transduction pathway that is involved in receptor phosphorylation remains elusive. This study provides evidence that the receptor of activated C kinase 1 (RACK1) and protein kinase C (PKC) signal transduction cascade is involved in the 20E-induced expression of the moult-associated transcription factor CHR3. A cDNA clone encoding a receptor of activated C kinase 1 was isolated from Choristoneura fumiferana (CfRACK1). This single copy gene coded a 36 kDa protein and was expressed ubiquitously in all of the developmental stages and the tissues tested, including the midgut, epidermis, fat body, head, Malpighian tubules, ovary and testis of larvae. High levels of the transcripts were also detected in a midgut-derived CF-203 cell line. We noticed that the green fluorescence protein-fused CfRACK1 protein was distributed in the cytosol surrounding the nuclei in stably transformed cells. Interference of CfRACK1 mRNA suppressed the 20E-induced expression of the transcription factor CHR3. Dequalinium-14; 1,1'-decamethylenebis-4-aminoquinaldinium diiodide (DECA), an inhibitor of RACK1 binding to protein kinase C, blocked the 20E-induced expression of CHR3 and accumulation of the ecdysone receptor (EcR) in the nuclei. All of these data together suggest that 20E-induced expression of CHR3 may involve phosphorylation of the ecdysone receptor component through the PKC/RACK1 signal transduction cascade, which facilitates the import of the receptor into the nuclei of cells.
Great Lakes Forestry Centre, Canadian Forest Service, Sault Ste. Marie, Ontario, Canada.
G X
GX
Quan
P J
PJ
Krell
B M
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Arif
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Feng
D021581:Active Transport, Cell Nucleus; D000595:Amino Acid Sequence; D000818:Animals; D003001:Cloning, Molecular; D004268:DNA-Binding Proteins / Q000096:biosynthesis / Q000235:genetics*; D004441:Ecdysterone / Q000502:physiology*; D018507:Gene Expression Regulation, Developmental; D019476:Insect Proteins / Q000096:biosynthesis* / Q000235:genetics; D008969:Molecular Sequence Data; D018978:Molting / Q000502:physiology; D009036:Moths / Q000235:genetics / Q000378:metabolism*; D010766:Phosphorylation; D011493:Protein Kinase C / Q000378:metabolism*; D021381:Protein Transport; D034622:RNA Interference; D000075370:Receptors for Activated C Kinase; D011956:Receptors, Cell Surface / Q000037:antagonists & inhibitors / Q000378:metabolism*; D018010:Receptors, Invertebrate Peptide / Q000096:biosynthesis / Q000235:genetics*; D011987:Receptors, Steroid; D017422:Sequence Analysis, DNA; D015398:Signal Transduction; D015534:Trans-Activators / Q000096:biosynthesis / Q000235:genetics*
C104701:CHR3 protein, Choristoneura fumiferana; D004268:DNA-Binding Proteins; D019476:Insect Proteins; D000075370:Receptors for Activated C Kinase; D011956:Receptors, Cell Surface; D018010:Receptors, Invertebrate Peptide; D011987:Receptors, Steroid; D015534:Trans-Activators; C055469:ecdysone receptor; D004441:Ecdysterone; D011493:Protein Kinase C