Molecular cloning of G proteins and phosphodiesterases from rat taste cells

• Published in: Physiology & behavior Vol. 56; no. 6; pp. 1157 - 1164
• Main Authors: McLaughlin, Susan K., McKinnon, Peter J., Spickofsky, Nancy, Danho, Waleed, Margolskee, Robert F.
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 01.12.1994
• Subjects: Amino Acid Sequence - genetics; Animals; cDNA; Cloning, Molecular; DNA, Complementary - genetics; GTP-Binding Proteins - genetics; Gustducin; Molecular Sequence Data; PCR taste transduction; Phosphoric Diester Hydrolases - genetics; Polymerase Chain Reaction; Rats; Rats, Sprague-Dawley; RNA; Signal Transduction - genetics; Synaptic Transmission - genetics; Taste - genetics; Taste Buds - physiology; Transducin; Transducin - genetics; Transducin; Gustducin; PCR taste transduction; cDNA; RNA
• ISSN: 0031-9384; 1873-507X
• DOI: 10.1016/0031-9384(94)90360-3

To identify and characterize those proteins involved in taste transduction, we cloned G proteins and phosphodiesterases from rat taste tissue. Using degenerate primers corresponding to conserved regions of G protein α subunits, the polymerase chain reaction was used to amplify and clone eight distinct cDNAs: α i-2, α i-3, α 12, α 14, α 8, α t-rod, α 1-cone and α gustducin. α i-3, α 14, α 8, and α t-rod are more highly expressed in taste tissue than in the surrounding nonsensory tissue. α gustducin is only expressed in taste cells. Rod transducin had previously been found only in the rod cells of the retina, where it converts light stimulation of rhodopsin into activation of cGMP phosphodiesterase. The primary sequence of α gustducin shows striking similarities to rod transducin in the receptor interaction domain and the phosphodiesterase activation site. We propose that gustducin and transducin regulate phosphodiesterase activity in taste cells and that this may promote bitter transduction and inhibit sweet transduction. Consistent with this proposal, we cloned two types of cAMP PDE from taste tissue: dnc-1 and PDE-3.