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J. Biol. Chem., Vol. 276, Issue 2, 1398-1406, January 12, 2001
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From the Department of Cellular and Molecular Medicine, University
of Ottawa, Ottawa, Ontario K1H 8M5, Canada
Novel splice variants of the
Alternative Splicing in Intracellular Loop Connecting Domains II
and III of the
1 Subunit of Cav1.2
Ca2+ Channels Predicts Two-domain Polypeptides with
Unique C-terminal Tails*
1 subunit of the Cav1.2 voltage-gated
Ca2+ channel were identified that predicted two truncated
forms of the 1 subunit comprising domains I and II
generated by alternative splicing in the intracellular loop region
linking domains II and III. In rabbit heart splice variant 1 (RH-1),
exon 19 was deleted, which resulted in a reading frameshift of exon 20 with a premature termination codon and a novel 19-amino acid
carboxyl-terminal tail. In the RH-2 variant, exons 17 and 18 were
deleted, leading to a reading frameshift of exons 19 and 20 with a
premature stop codon and a novel 62-amino acid carboxyl-terminal tail.
RNase protection assays with RH-1 and RH-2 cRNA probes confirmed the expression in cardiac and neuronal tissue but not skeletal muscle. The
deduced amino acid sequence from full-length cDNAs encoding the two
variants predicted polypeptides of 99.0 and 99.2 kDa, which constituted
domains I and II of the 1 subunit of the
Cav1.2 channel. Antipeptide antibodies directed to
sequences in the second intracellular loop between domains II and
III identified the 240-kDa Cav1.2 subunit in
sarcolemmal and heavy sarcoplasmic reticulum (HSR) membranes and a
99-kDa polypeptide in the HSR. An antipeptide antibody raised against
unique sequences in the RH-2 variant also identified a 99-kDa
polypeptide in the HSR. These data reveal the expression of additional
Ca2+ channel structural units generated by alternative
splicing of the Cav1.2 gene.
*
The costs of publication of this
article were defrayed in part by the
payment of page charges. The article
must therefore be hereby marked
"advertisement" in
accordance with 18 U.S.C. Section
1734 solely to indicate this fact.
Recipient of a grant from the Medical Research Council of Canada
and Career Investigator of the Heart and Stroke Foundation of Ontario.
To whom correspondence should be addressed: Dept. of Cellular and
Molecular Medicine, University of Ottawa, 451 Smyth Rd., Ottawa,
Ontario, Canada K1H 8H5. Phone: 613-562-5800 (ext. 8355); Fax:
613-562-5434.
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