Elsevier

Genomics

Volume 61, Issue 2, 15 October 1999, Pages 201-209
Genomics

Regular Article
Genomic Structure and Functional Expression of a Human α2/δ Calcium Channel Subunit Gene (CACNA2)

https://doi.org/10.1006/geno.1999.5941Get rights and content

Abstract

CACNA2 encodes the α2/δ subunit of the human voltage-gated calcium channels and is located in the candidate region of malignant hyperthermia susceptibility type 3 (MHS3). We determined the structural organization of CACNA2 by isolation of overlapping genomic DNA clones from a human phage library. The gene consists of at least 40 exons, 2 of which are alternatively spliced, spanning more than 150 kb of genomic DNA. Exons range from 21 to 159 bp, and introns range from 98 bp to at least more than 20 kb. We constructed a full-length cDNA and cloned it into a mammalian expression vector. Cotransfection of the CACNA2 cDNA with α1A and β4 cDNA into HEK293 cells led to the expression of Q-type calcium currents. The α2/δ subunit enhanced the current density 18-fold compared to cells transfected with only α1A and β4 cDNA. The sequence analysis provides the basis for comprehensive mutation screening of CACNA2 for putative MHS3 individuals and patients with other channelopathies.

References (38)

  • L.J. Ptacek et al.

    Dihydropyridine receptor mutations cause hypokalemic periodic paralysis

    Cell

    (1994)
  • M.E. Williams et al.

    Structure and functional expression of alpha 1, alpha 2, and beta subunits of a novel human neuronal calcium channel subtype

    Neuron

    (1992)
  • R. Bangalore et al.

    Influence of L-type Ca channel alpha 2/delta-subunit on ionic and gating current in transiently transfected HEK 293 cells

    Am. J. Physiol.

    (1996)
  • M.J. Berridge

    Elementary and global aspects of calcium signalling

    J. Physiol.

    (1997)
  • M. De Waard et al.

    Subunit regulation of the neuronal alpha 1A Ca2+ channel expressed in Xenopus oocytes

    J. Physiol.

    (1995)
  • S.J. Elledge et al.

    Lambda YES: A multifunctional cDNA expression vector for the isolation of genes by complementation of yeast and Escherichia coli mutations

    Proc. Natl. Acad. Sci. USA

    (1991)
  • S.B. Ellis et al.

    Sequence and expression of mRNAs encoding the alpha 1 and alpha 2 subunits of a DHP-sensitive calcium channel

    Science

    (1988)
  • R. Felix et al.

    Dissection of functional domains of the voltage-dependent Ca2+ channel alpha2delta subunit

    J. Neurosci.

    (1997)
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      HEK-293 and tsA-201 cells were transfected using standard calcium phosphate method. For whole-cell and single-channel recordings, HEK-293 cells were transfected with a 1: 0.5: 1.5 ratio of CaV1.2 (α1c77; Soldatov et al., 1995), either human WT or mutant β2d-subunit and human α2δ1-subunit (Schleithoff et al., 1999). For co-immunoprecipitation experiments, tsA-201 cells were transfected with a 1:2 ratio of either human WT or mutant β2d_HA-subunit and human Flag-Gem.

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    Sequence data from this article have been deposited with the EMBL/GenBank Data Libraries under Accession Nos. AF083817–AF083854.

    1

    To whom correspondence should be addressed at Department of Applied Physiology, University of Ulm, Albert-Einstein-Allee 11, D-89069 Ulm, Germany. Telephone: ++49-731-502-3250. Fax: ++49-731-502-3260. E-mail: [email protected].

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