Molecular cloning of G proteins and phosphodiesterases from rat taste cells.
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 alpha subunits, the polymerase chain reaction was used to amplify and clone eight distinct cDNAs: alpha i-2, alpha i-3, alpha 12, alpha 14, a(s), alpha t-rod, alpha t-cone and alpha gustducin. alpha i-3, alpha 14, alpha s, and alpha t-rod are more highly expressed in taste tissue than in the surrounding nonsensory tissue. alpha 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 alpha 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.
McLaughlin
S K
SK
Roche Research Center, Roche Institute of Molecular Biology, Nutley, NJ 07110.
McKinnon
P J
PJ
Spickofsky
N
N
Danho
W
W
Margolskee
R F
RF
D000595:Amino Acid Sequence / Q000235:genetics; D000818:Animals; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000235:genetics; D019204:GTP-Binding Proteins / Q000235:genetics*; D008969:Molecular Sequence Data; D010727:Phosphoric Diester Hydrolases / Q000235:genetics*; D016133:Polymerase Chain Reaction; D051381:Rats; D017207:Rats, Sprague-Dawley; D015398:Signal Transduction / Q000235:genetics; D009435:Synaptic Transmission / Q000235:genetics*; D013649:Taste / Q000235:genetics*; D013650:Taste Buds / Q000502:physiology*; D015293:Transducin / Q000235:genetics
D018076:DNA, Complementary; C075192:gustducin; D010727:Phosphoric Diester Hydrolases; D019204:GTP-Binding Proteins; D015293:Transducin