Goldin, Alan L. and Snutch, Terry P. and Lübbert, Hermann and Dowsett, Andrew and Marshall, John and Auld, Vanessa and Downey, William and Fritz, Larry C. and Lester, Henry A. and Dunn, Robert and Catterall, William A. and Davidson, Norman (1986) Messenger RNA coding for only the alpha subunit of the rat brain Na channel is sufficient for expression of functional channels in Xenopus oocytes. Proceedings of the National Academy of Sciences of the United States of America, 83 (19). pp. 7503-7507. ISSN 0027-8424 http://resolver.caltech.edu/CaltechAUTHORS:GOLpnas86
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Several cDNA clones coding for the high molecular weight (alpha) subunit of the voltage-sensitive Na channel have been selected by immunoscreening a rat brain cDNA library constructed in the expression vector lambda gt11. As will be reported elsewhere, the amino acid sequence translated from the DNA sequence shows considerable homology to that reported for the Electrophorus electricus electroplax Na channel. Several of the cDNA inserts hybridized with a low-abundance 9-kilobase RNA species from rat brain, muscle, and heart. Sucrose-gradient fractionation of rat brain poly(A) RNA yielded a high molecular weight fraction containing this mRNA, which resulted in functional Na channels when injected into oocytes. This fraction contained undetectable amounts of low molecular weight RNA. The high molecular weight Na channel RNA was selected from rat brain poly(A) RNA by hybridization to a single-strand antisense cDNA clone. Translation of this RNA in Xenopus oocytes resulted in the appearance of tetrodotoxin-sensitive voltage-sensitive Na channels in the oocyte membrane. These results demonstrate that mRNA encoding the alpha subunit of the rat brain Na channel, in the absence of any beta-subunit mRNA, is sufficient for translation to give functional channels in oocytes.
|Additional Information:||© 1986 by the National Academy of Sciences. Contributed by Norman Davidson, October 28, 1985. We thank Nathan Dascal for his expert assistance and advice in performing the electrophysiological procedures. This research has been supported by grants from the National Institutes of Health to N.D., W.A.C., and H.A.L.; by a grant from the Medical Research Council of Canada to R.D.; and by fellowship support from the American Heart Association to A.D. and T.S.; from the Natural Sciences and Engineering Research Council of Canada to T.S.; from the Deutsche Forschungsgemeinschaft to H.L.; and from the National Multiple Sclerosis Society to A.L.G. The publication costs of this article were defrayed in part by page charge payment. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. §1734 solely to indicate this fact.|
|Subject Keywords:||hybrid selection; sucrose-gradient fractionation; voltage clamp|
|Usage Policy:||No commercial reproduction, distribution, display or performance rights in this work are provided.|
|Deposited By:||Tony Diaz|
|Deposited On:||09 Oct 2008 15:36|
|Last Modified:||09 Jan 2013 18:09|
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