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Reconstitution of mutant V2 vasopressin receptors by adenovirus mediated gene transfer. Molecular basis and clinical implication

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Item Type:Article
Title:Reconstitution of mutant V2 vasopressin receptors by adenovirus mediated gene transfer. Molecular basis and clinical implication
Creators Name:Schoeneberg, T. and Sandig, V. and Wess, J. and Gudermann, T. and Schultz, G.
Abstract:Recent studies with transfected COS-7 cells have shown that functionally inactive mutant V2 vasopressin receptors (occurring in patients with nephrogenic diabetes insipidus) can be functionally rescued by coexpression of a carboxy-terminal V2 receptor fragment (V2-tail) spanning the region where various mutations occur [Schoneberg, T., J. Yun, D. Wenkert, and J. Wess. 1996. EMBO (Eur. Mol. Biol. Organ.) J. 15:1283-1291]. In this study, we set out to characterize the underlying molecular mechanism. Using a coimmunoprecipitation strategy and a newly developed sandwich ELISA system, a direct and highly specific interaction between the mutant V2 vasopressin receptor proteins and the V2-tail polypeptide was demonstrated. To study the potential therapeutic usefulness of these findings, Chinese hamster ovary (CHO) cell lines stably expressing low levels of functionally inactive mutant V2 vasopressin receptors were created and infected with a recombinant adenovirus carrying the V2-tail gene fragment. After adenovirus infection, vasopressin gained the ability to stimulate cAMP formation with high potency and efficacy in all CHO cell clones studied. Moreover, adenovirus-mediated gene transfer also proved to be a highly efficient method for achieving expression of the V2-tail fragment (as well as the wild-type V2 receptor) in Madin-Darby canine kidney tubular cells. Taken together, these studies clarify the molecular mechanisms by which receptor fragments can restore function of mutationally inactivated G protein-coupled receptors and suggest that adenovirus-mediated expression of receptor fragments may lead to novel strategies for the treatment of a variety of human diseases.
Keywords:Nephrogenic Diabetes Insipidus, Vasopressin Receptor, Gene Therapy, Receptor Mutagenesis, Receptor Folding, Animals, Cricetinae, Dogs
Source:Journal of Clinical Investigation
ISSN:0021-9738
Publisher:American Society for Clinical Investigation (U.S.A.)
Volume:100
Number:6
Page Range:1547-1556
Date:15 September 1997
Official Publication:https://doi.org/10.1172/JCI119678
PubMed:View item in PubMed

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