A spatiotemporally coordinated cascade of protein kinase C activation controls isoform-selective translocation.
In pituitary GH3B6 cells, signaling involving the protein kinase C (PKC) multigene family can self-organize into a spatiotemporally coordinated cascade of isoform activation. Indeed, thyrotropin-releasing hormone (TRH) receptor activation sequentially activated green fluorescent protein (GFP)-tagged or endogenous PKCbeta1, PKCalpha, PKCepsilon, and PKCdelta, resulting in their accumulation at the entire plasma membrane (PKCbeta and -delta) or selectively at the cell-cell contacts (PKCalpha and -epsilon). The duration of activation ranged from 20 s for PKCalpha to 20 min for PKCepsilon. PKCalpha and -epsilon selective localization was lost in the presence of Gö6976, suggesting that accumulation at cell-cell contacts is dependent on the activity of a conventional PKC. Constitutively active, dominant-negative PKCs and small interfering RNAs showed that PKCalpha localization is controlled by PKCbeta1 activity and is calcium independent, while PKCepsilon localization is dependent on PKCalpha activity. PKCdelta was independent of the cascade linking PKCbeta1, -alpha, and -epsilon. Furthermore, PKCalpha, but not PKCepsilon, is involved in the TRH-induced beta-catenin relocation at cell-cell contacts, suggesting that PKCepsilon is not the unique functional effector of the cascade. Thus, TRH receptor activation results in PKCbeta1 activation, which in turn initiates a calcium-independent but PKCbeta1 activity-dependent sequential translocation of PKCalpha and -epsilon. These results challenge the current understanding of PKC signaling and raise the question of a functional dependence between isoforms.
Collazos
Alejandra
A
Institut de Génomique Fonctionnelle, 141 rue de la Cardonille, F-34094 Montpellier Cedex 5, France.
Diouf
Barthélémy
B
Guérineau
Nathalie C
NC
Quittau-Prévostel
Corinne
C
Peter
Marion
M
Coudane
Fanny
F
Hollande
Frédéric
F
Joubert
Dominique
D
D000818:Animals; D002118:Calcium / Q000378:metabolism; D002227:Carbazoles / Q000494:pharmacology; D002450:Cell Communication; D002460:Cell Line; D004789:Enzyme Activation; D004791:Enzyme Inhibitors / Q000494:pharmacology; D007211:Indoles / Q000494:pharmacology; D015759:Ionomycin / Q000494:pharmacology; D007527:Isoenzymes / Q000235:genetics / Q000378:metabolism; D010902:Pituitary Gland / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D011493:Protein Kinase C / Q000037:antagonists & inhibitors / Q000187:drug effects / Q000235:genetics / Q000378:metabolism*; D064546:Protein Kinase C beta; D021381:Protein Transport; D051381:Rats; D018025:Receptors, Thyrotropin-Releasing Hormone / Q000187:drug effects / Q000378:metabolism; D015398:Signal Transduction; D013973:Thyrotropin-Releasing Hormone / Q000378:metabolism / Q000494:pharmacology; D051176:beta Catenin / Q000187:drug effects / Q000378:metabolism
D002227:Carbazoles; D004791:Enzyme Inhibitors; D007211:Indoles; D007527:Isoenzymes; D018025:Receptors, Thyrotropin-Releasing Hormone; D051176:beta Catenin; C081021:Go 6976; D015759:Ionomycin; D013973:Thyrotropin-Releasing Hormone; D011493:Protein Kinase C; D064546:Protein Kinase C beta; D002118:Calcium