Prenylation of mammalian Ras protein in Xenopus oocytes.
Ras protein requires an intermediate of the cholesterol biosynthetic pathway for posttranslational modification and membrane anchorage. This step is necessary for biological activity. Maturation of Xenopus laevis oocytes induced by an oncogenic human Ras protein can be inhibited by lovastatin or compactin, inhibitors of the synthesis of mevalonate, an intermediate of cholesterol biosynthesis. This inhibition can be overcome by mevalonic acid or farnesyl diphosphate, a cholesterol biosynthetic intermediate downstream of mevalonate, but not by squalene, an intermediate after farnesyl pyrophosphate in the pathway. This study supports the idea that in Xenopus oocytes, the Ras protein is modified by a farnesyl moiety or its derivative. Furthermore, an octapeptide with the sequence similar to the C-terminus of the c-H-ras protein inhibits the biological activity of Ras proteins in vivo, suggesting that it competes for the enzyme or enzymes responsible for transferring the isoprenoid moiety (prenylation) in the oocytes. This inhibition of Ras prenylation by the peptide was also observed in vitro, using both Saccharomyces cerevisiae and Xenopus oocyte extracts. These observations show that Xenopus oocytes provide a convenient in vivo system for studies of inhibitors of the posttranslational modification of the Ras protein, especially for inhibitors such as peptides that do not penetrate cell membranes.
Kim
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Lawrence Berkeley Laboratory, University of California, Berkeley 94720.
Rine
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Kim
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D000595:Amino Acid Sequence; D000818:Animals; D002784:Cholesterol / Q000096:biosynthesis; D005260:Female; D005813:Genes, Synthetic; D008798:Mevalonic Acid / Q000378:metabolism; D008845:Microinjections; D008969:Molecular Sequence Data; D009842:Oligopeptides / Q000138:chemical synthesis / Q000494:pharmacology; D015689:Oncogene Protein p21(ras) / Q000235:genetics / Q000378:metabolism*; D009865:Oocytes / Q000378:metabolism*; D011106:Polyisoprenyl Phosphates / Q000378:metabolism*; D011499:Protein Processing, Post-Translational / Q000187:drug effects; D012717:Sesquiterpenes; D014162:Transfection; D014982:Xenopus laevis
D009842:Oligopeptides; D011106:Polyisoprenyl Phosphates; D012717:Sesquiterpenes; C004808:farnesyl pyrophosphate; D002784:Cholesterol; D015689:Oncogene Protein p21(ras); D008798:Mevalonic Acid