The gamma subunit of transducin is farnesylated.
Protein prenylation with farnesyl or geranylgeranyl moieties is an important posttranslational modification that affects the activity of such diverse proteins as the nuclear lamins, the yeast mating factor mata, and the ras oncogene products. In this article, we show that whole retinal cultures incorporate radioactive mevalonic acid into proteins of 23-26 kDa and one of 8 kDa. The former proteins are probably the "small" guanine nucleotide-binding regulatory proteins (G proteins) and the 8-kDa protein is the gamma subunit of the well-studied retinal heterotrimeric G protein (transducin). After deprenylating purified transducin and its subunits with Raney nickel or methyl iodide/base, the adducted prenyl group can be identified as an all-trans-farnesyl moiety covalently linked to a cysteine residue. Thus far, prenylation reactions have been found to occur at cysteine in a carboxyl-terminal consensus CAAX sequence, where C is the cysteine, A is an aliphatic amino acid, and X is undefined. Both the alpha and gamma subunits of transducin have this consensus sequence, but only the gamma subunit is prenylated. Therefore, the CAAX motif is not necessary and sufficient to direct prenylation. Finally, since transducin is the best understood G protein, both structurally and mechanistically, the discovery that it is farnesylated should allow for a quantitative understanding of this post-translational modification.
Lai
R K
RK
Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Boston, MA 02115.
Perez-Sala
D
D
Cañada
F J
FJ
Rando
R R
RR
D000818:Animals; D002417:Cattle; D004224:Diterpenes / Q000032:analysis*; D005204:Farnesol / Q000032:analysis*; D008401:Gas Chromatography-Mass Spectrometry; D066298:In Vitro Techniques; D007930:Leucine / Q000378:metabolism; D046911:Macromolecular Substances; D008798:Mevalonic Acid / Q000378:metabolism; D011499:Protein Processing, Post-Translational; D012160:Retina / Q000378:metabolism*; D015293:Transducin / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics
D004224:Diterpenes; D046911:Macromolecular Substances; D005204:Farnesol; C017338:geranylgeraniol; D015293:Transducin; D007930:Leucine; D008798:Mevalonic Acid