Depolarization regulates cyclin D1 degradation and neuronal apoptosis: a hypothesis about the role of the ubiquitin/proteasome signalling pathway.
Depolarization and subsequent calcium entry exert essential neuroprotective effects but the ultimate effector by which calcium blocks apoptosis is not known. Here we show that inhibition of calcium entry into cerebellar neurons by switching from high to low extracellular K+ concentrations (30-5 mM) induces apoptosis, that correlates with a rapid accumulation of cyclin D1 (CD1), an early marker of the G1/S transition of the cell cycle. These effects on apoptosis and cyclin D1 are mimicked either by blocking calcium entry into neurons (LaCl3, 100 microM or nifedipine, 10(-6) M) or by inhibiting the calcium/calmodulin pathway (calmidazolium, 10(-7) M). The increased CD1 protein levels do not result from a transcriptional upregulation of the CD1 gene by the Ca2+/calmodulin pathway but rather reflect an accumulation due to the lack of degradation by the proteasome-dependent pathway. Specific proteasome antagonists: carbobenzoxyl-leucinyl-leucinyl-norvalinal-H (MG-115), carbobenzoxyl-leucinyl-leucinyl-leucinal-H (MG-132) and clastolactacystin beta-lactone, induce neuronal apoptosis by themselves. Finally, this pathway is functional only at neuroprotective concentrations of K+ (30 mM), suggesting that calcium/CamK signalling pathway may regulate neuronal death by regulating the proteasome-mediated degradation activity of rapidly turning-over proteins (constitutively expressed genes or pre-existing pools of mRNA).
UMR 7519 CNRS, IPCB, Strasbourg, France.
A L
AL
Boutillier
P
P
Kienlen-Campard
J P
JP
Loeffler
D000818:Animals; D017209:Apoptosis / Q000502:physiology*; D002118:Calcium / Q000502:physiology; D002147:Calmodulin / Q000502:physiology; D002531:Cerebellum / Q000166:cytology; D017868:Cyclic AMP-Dependent Protein Kinases / Q000037:antagonists & inhibitors / Q000378:metabolism; D019938:Cyclin D1 / Q000235:genetics / Q000378:metabolism*; D003546:Cysteine Endopeptidases / Q000378:metabolism*; D015853:Cysteine Proteinase Inhibitors / Q000494:pharmacology; D017931:DNA Primers; D004791:Enzyme Inhibitors / Q000494:pharmacology; D015870:Gene Expression / Q000502:physiology; D007093:Imidazoles / Q000494:pharmacology; D020287:In Situ Nick-End Labeling; D007546:Isoquinolines / Q000494:pharmacology; D007976:Leupeptins / Q000494:pharmacology; D008564:Membrane Potentials / Q000187:drug effects / Q000502:physiology; D051379:Mice; D008815:Mice, Inbred Strains; D009097:Multienzyme Complexes / Q000378:metabolism*; D009474:Neurons / Q000166:cytology* / Q000201:enzymology; D011189:Potassium Chloride / Q000494:pharmacology; D011480:Protease Inhibitors / Q000494:pharmacology; D046988:Proteasome Endopeptidase Complex; D012333:RNA, Messenger / Q000032:analysis; D015398:Signal Transduction / Q000502:physiology; D013449:Sulfonamides; D014452:Ubiquitins / Q000378:metabolism*
D002147:Calmodulin; D015853:Cysteine Proteinase Inhibitors; D017931:DNA Primers; D004791:Enzyme Inhibitors; D007093:Imidazoles; D007546:Isoquinolines; D007976:Leupeptins; D009097:Multienzyme Complexes; D011480:Protease Inhibitors; D012333:RNA, Messenger; D013449:Sulfonamides; D014452:Ubiquitins; C089486:carbobenzoxy-leucyl-leucyl-norvalinal; D019938:Cyclin D1; C031938:calmidazolium; D011189:Potassium Chloride; D017868:Cyclic AMP-Dependent Protein Kinases; D003546:Cysteine Endopeptidases; D046988:Proteasome Endopeptidase Complex; C063509:N-(2-(4-bromocinnamylamino)ethyl)-5-isoquinolinesulfonamide; C072553:benzyloxycarbonylleucyl-leucyl-leucine aldehyde; D002118:Calcium