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The Journal of Neuroscience, April 15, 2003, 23(8):3446
Interactions with PDZ Proteins Are Required for L-Type Calcium
Channels to Activate cAMP Response Element-Binding
Protein-Dependent Gene Expression
Jason P.
Weick,
Rachel D.
Groth,
Ann L.
Isaksen, and
Paul G.
Mermelstein
Department of Neuroscience, University of Minnesota, Minneapolis,
Minnesota 55455
After brief periods of heightened stimulation, calcium
entry through L-type calcium channels leads to activation of the
transcription factor cAMP response element-binding protein (CREB) and
CRE-dependent transcription. Many of the details surrounding the
mechanism by which L-type calcium channels are privileged in signaling
to CREB, to the exclusion of other calcium entry pathways, has remained unclear. We hypothesized that the PDZ interaction sequence
contained within the last four amino acids of the calcium channel
1C (CaV1.2) subunit [Val-Ser-Asn-Leu
(VSNL)] is critical for L-type calcium channels (LTCs) to interact
with the signaling machinery that triggers activity-dependent gene
expression. To disrupt this interaction, hippocampal CA3-CA1 pyramidal
neurons were transfected with DNA encoding for enhanced green
fluorescent protein tethered to VSNL (EGFP-VSNL). EGFP-VSNL
significantly attenuated L-type calcium channel-induced CREB
phosphorylation and CRE-dependent transcription, although somatic
calcium concentrations after stimulation remained unchanged. The effect
of EGFP-VSNL was specific to the actions of L-type calcium channels,
because CREB signaling after NMDA receptor stimulation remained intact.
The importance of the PDZ interaction sequence was verified using
dihydropyridine (DHP)-insensitive 1C subunits.
Neurons transfected with 1C lacking the terminal five
amino acids (DHP-LTCnoPDZ) exhibited attenuated CREB responses in
comparison with cells expressing the full-length subunit (DHP-LTC). Collectively, these data suggest that localized calcium responses, regulated by interactions with PDZ domain proteins, are necessary for
L-type calcium channels to effectively activate CREB and CRE-mediated gene expression.
Key words:
NIL-16; CREB; NFAT; L-type calcium channel; 1C; CaV1.2; CIPP; NMDA
Copyright © 2003 Society for Neuroscience 0270-6474/03/2383446-11$05.00/0
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