Classical and novel protein kinase C (PKC) isoforms are down-regulated as a result of chronic activation by certain tumour promoters and physiological stimuli; however, the mechanisms leading to down-regulation are not fully understood. In the present study, we have studied the PMA ('TPA')-induced degradation of PKCΔ in NIH 3T3 cells under culture conditions where PKCΔ displays cell-cycle-dependent down-regulation. In contrast with previous studies, a hyperphosphorylated form of this PKC isoform, promoted by calyculin A, was rapidly degraded in PMA-treated cells. Similarly, the presence of calyculin A enhanced the down-regulation of PKCΔ observed on G1/S-phase progression through the cell cycle. Analysis of phosphorylation-site mutants indicated that the T-loop Thr505 phosphorylation site was critical for induced degradation.
- protein phosphatase
↵1 Present address: CNRS UMR 144, Institut Curie, 26, rue d'Ulm, 75248 Paris Cedex 05, France.
Abbreviations used: BIM-I, bisindolylmaleimide I; FCS, foetal calf serum; GFP, green fluorescent protein; HA, haemagglutinin; MEF, mouse embryonic fibroblast; PKC, protein kinase C; PP1, protein phosphatase 1; PP2A, protein phosphatase 2A; T505A (etc.), Thr505→Ala mutant.
- The Biochemical Society, London ©2002