Abstract
The L-type voltage-gated calcium channel Cav1.2 and the calcium-activated CaM kinase cascade both regulate excitation transcription coupling in the brain. CaM kinase is known to associate with the C terminus of Cav1.2 in a region called the PreIQ-IQ domain, which also binds multiple calmodulin molecules. Here we identify and characterize a second CaMKII binding site in the N terminus of Cav1.2 that is formed by a stretch of four amino residues (cysteine–isoleucine–serine–isoleucine) and which regulates channel expression and function. By using live cell imaging of tsA-201 cells we show that GFP fusion constructs of the CaMKII binding region, termed N2B-II co-localize with mCherry-CaMKII. Mutating CISI to AAAA ablates binding to and colocalization with CaMKII. Cav1.2-AAAA channels show reduced cell surface expression in tsA-201 cells, but interestingly, display an increase in channel function that offsets the trafficking deficit. Altogether our data reveal that the proximal N terminus of Cav1.2 contains a CaMKII binding region which contributes to channel surface expression and function.
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Abbreviations
- CaMK:
-
Calmodulin kinase
- CaMKK:
-
Calmodulin kinase kinase
- CaM:
-
Calmodulin
- CDF:
-
Calcium-dependent facilitation
- CDI:
-
Calcium-dependent inactivation
- VDF:
-
Voltage-dependent facilitation
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Acknowledgments
This work was supported by a grant from the Natural Sciences and Engineering Research Council. BAS is supported by a studentship from Alberta Innovates-Health Solutions (AI-HS). IAS is supported by a postdoctoral fellowship from Mitacs Elevate. GWZ is an AI-HS Scientist and a Canada Research Chair.
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The authors declare that they have no conflict of interest.
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All experiments performed in this manuscript comply with the laws of Canada.
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B.A. Simms and I.A. Souza contributed equally to this study.
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Simms, B.A., Souza, I.A., Rehak, R. et al. The Cav1.2 N terminus contains a CaM kinase site that modulates channel trafficking and function. Pflugers Arch - Eur J Physiol 467, 677–686 (2015). https://doi.org/10.1007/s00424-014-1538-7
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DOI: https://doi.org/10.1007/s00424-014-1538-7