Novel Down-Regulatory Mechanism of the Surface Expression of Vasopressin V2 Receptor by an Alternative Splice Receptor Variant

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Title: Novel Down-Regulatory Mechanism of the Surface Expression of Vasopressin V2 Receptor by an Alternative Splice Receptor Variant
Authors: Sarmiento, Jose M.; Anazco, Carolina C.; Campos, Danae M.; Prado, Gregory N.; Navarro, Javier; Gonzalez, Carlos B.
Publisher: Journal of Biological Chemistry
Date Published: September 08, 2004
Reference Number: 661
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In rat kidney, two alternatively spliced transcripts are generated from the V( 2) vasopressin receptor gene. The large transcript (1.2 kb) encodes the canonical V( 2) receptor, whereas the small transcript encodes a splice variant displaying a distinct sequence corresponding to the putative 7( th) transmembrane domain and the intracellular C-terminus of V( 2) receptor. This work showed that the small spliced transcript is translated in the rat kidney collecting tubules. However, the protein encoded by the small transcript (here called V( 2b) splice variant) is retained inside the cell, in contrast to the preferential surface distribution of the V( 2) receptor (here called V( 2a) receptor). Cells expressing the V( 2b) splice variant do not exhibit binding to [(3)H] labelled vasopressin. Interestingly, we found that expression of the splice variant V( 2b) down-regulates the surface expression of V( 2a) receptor, most likely via the formation V( 2a).V( 2b) heterodimers, as demonstrated by co-immunoprecipitation and fluorescence resonance energy transfer experiments between V( 2a) receptor and V( 2b) splice variant. The V( 2b) splice variant would be then acting as a dominant negative. The effect of V( 2b) splice variant is specific, as it does not affect the surface expression of the G protein-coupled IL-8 receptor (CXCR1). Furthermore, the sequence encompassing residues 242-339 corresponding to the C-terminal of the V( 2b) splice variant, also down-regulates the surface expression of the V( 2a) receptor. We suggest that some forms of nephrogenic diabetes insipidus are due to the overexpression of the splice variant V( 2b), which could retain the wild-type V( 2a) receptor inside the cell via the formation of V( 2a)*V( 2b) heterodimers.
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