A carboxyl-terminal cysteine residue is required for palmitic acid binding and biological activity of the ras-related yeast YPT1 protein.
The Saccharomyces cerevisiae YPT1 gene codes for a ras-like, guanine nucleotide-binding protein which is essential for cell viability. The functional significance of two consecutive cysteines at the very carboxyl-terminal end of this protein and in ypt homologues of other eukaryotic species was examined. YPT1 gene mutations were generated that either led to substitutions by serine or the deletion of one or both C-terminal cysteines. The consequences of the mutations were checked in cells after replacing the wild type with the mutant genes. It was found that as long as one of the cysteines was retained, the protein was fully functional. The YPT1 protein could be labelled with [3H]palmitic acid that appeared to be bound in an ester linkage. The wild-type protein was evenly distributed between soluble and membrane-associated proteins, the palmitoylated form was predominantly in the crude membrane fraction. The mutant protein lacking the C-terminal cysteines was not palmitoylated and was exclusively found in the soluble fraction. The extension by three residues, -Val-Leu-Ser, generating a ras-typical C-terminal end, did not interfere with the mutant YPT1 protein's function although it resulted in a reduced labelling with palmitic acid.
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D003062:Codon; D003545:Cysteine; D005656:Fungal Proteins / Q000235:genetics* / Q000378:metabolism; D005796:Genes; D005800:Genes, Fungal; D006801:Humans; D008969:Molecular Sequence Data; D009154:Mutation; D019308:Palmitic Acid; D010169:Palmitic Acids / Q000378:metabolism*; D011485:Protein Binding; D012441:Saccharomyces cerevisiae / Q000235:genetics* / Q000378:metabolism; D012689:Sequence Homology, Nucleic Acid; D018631:ras Proteins
D003062:Codon; D005656:Fungal Proteins; D010169:Palmitic Acids; D019308:Palmitic Acid; D018631:ras Proteins; D003545:Cysteine