Expression of cone transducin, Gz alpha, and other G-protein alpha-subunit messenger ribonucleic acids in pancreatic islets.
The G-proteins are a family of heterotrimeric guanine nucleotide-binding proteins that play important roles in signal transduction and whose expression is regulated in a tissue-specific manner. Here we have surveyed the expression of G-protein alpha-subunits in mouse pancreatic islets. Degenerate oligonucleotide primers corresponding to conserved primary sequences in known G alpha-subunits were used in a reverse transcriptase-polymerase chain reaction, and the amplified complementary DNA (cDNA) fragments were subcloned and sequenced. Over 100 clones were analyzed, from which we determined that islet cells express at least seven G alpha-subunits: G8 alpha, Gi1 alpha or Gi3 alpha, Gi2 alpha, G11 alpha, G14 alpha, Gz alpha, and Gt2 alpha (cone transducin). In particular, the identification of Gz alpha and Gt2 alpha was of interest in that previous studies had indicated that the expression of Gz alpha was restricted mainly to the brain, retina, and adrenal gland, whereas Gt2 alpha was expressed predominantly in retinal cone photoreceptors. By Western blot analysis, we estimated that the amount of Gz alpha protein present in mouse islets was about 40% of that in retina. To further investigate the expression of Gt2 alpha, mouse Gt2 alpha cDNA was cloned from a retinal library and sequenced. The cDNA was used as a probe for Northern blot analysis, and the results confirmed that mouse islets contained a substantial level of Gt2 alpha messenger RNA (mRNA), albeit less than that found in retina (approximately 5-fold lower). Gt2 alpha mRNA was also shown to be present in a clonal mouse pancreatic alpha-cell line (alpha TC1-6) as well as in adrenal gland, pituitary, and a clonal mouse anterior pituitary cell line (AtT20). In situ hybridization revealed that Gt2 alpha mRNA was expressed essentially throughout the islet, suggesting that it is normally expressed in the abundant islet beta-cells and possibly others. In situ analysis also showed that Gt2 alpha mRNA expressed in the pituitary was limited to the intermediate and anterior lobes. We conclude that islet cells express multiple G-proteins, including several that are normally expressed at high levels in certain neuronal cells.
Department of Biochemistry and Molecular Biology, University of Chicago, Illinois 60637.
J M
JM
Zigman
G T
GT
Westermark
J
J
LaMendola
D F
DF
Steiner
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D019204:GTP-Binding Proteins / Q000235:genetics*; D007515:Islets of Langerhans / Q000378:metabolism*; D051379:Mice; D008969:Molecular Sequence Data; D015345:Oligonucleotide Probes / Q000235:genetics; D016133:Polymerase Chain Reaction; D012333:RNA, Messenger / Q000378:metabolism*; D017949:Retinal Cone Photoreceptor Cells / Q000378:metabolism*; D014158:Transcription, Genetic; D015293:Transducin / Q000235:genetics*
D015345:Oligonucleotide Probes; D012333:RNA, Messenger; D019204:GTP-Binding Proteins; D015293:Transducin