N-terminally myristoylated Ras proteins require palmitoylation or a polybasic domain for plasma membrane localization

Autor: H. Paterson, John F. Hancock, K. A. Cadwallader, S. G. Macdonald
Rok vydání: 1994
Předmět:
Signal peptide
Molecular Sequence Data
Restriction Mapping
Palmitic Acid
Palmitic Acids
Biology
Transfection
Myristic Acid
Polymerase Chain Reaction
Cell Line
Proto-Oncogene Proteins p21(ras)
Cell membrane
Palmitoylation
Prenylation
Mutant protein
Chlorocebus aethiops
Serine
medicine
Animals
Point Mutation
Amino Acid Sequence
Cysteine
Protein kinase A
Molecular Biology
DNA Primers
Myristoylation
Mitogen-Activated Protein Kinase Kinases
Base Sequence
Sequence Homology
Amino Acid

Cell Membrane
Cell Biology
Recombinant Proteins
Cell biology
medicine.anatomical_structure
Biochemistry
Protein Biosynthesis
Calcium-Calmodulin-Dependent Protein Kinases
Mutagenesis
Site-Directed

lipids (amino acids
peptides
and proteins)

Myristic Acids
Protein Kinases
Protein Processing
Post-Translational

Research Article
Zdroj: Molecular and Cellular Biology. 14:4722-4730
ISSN: 1098-5549
0270-7306
DOI: 10.1128/mcb.14.7.4722
Popis: Plasma membrane targeting of Ras requires CAAX motif modifications together with a second signal from an adjacent polybasic domain or nearby cysteine palmitoylation sites. N-terminal myristoylation is known to restore membrane binding to H-ras C186S (C-186 is changed to S), a mutant protein in which all CAAX processing is abolished. We show here that myristoylated H-ras C186S is a substrate for palmitoyltransferase, despite the absence of C-terminal farnesylation, and that palmitoylation is absolutely required for plasma membrane targeting of myristoylated H-ras. Similarly, the polybasic domain is required for specific plasma membrane targeting of myristoylated K-ras. In contrast, the combination of myristoylation plus farnesylation results in the mislocalization of Ras to numerous intracellular membranes. Ras that is only myristoylated does not bind with a high affinity to any membrane. The specific targeting of Ras to the plasma membrane is therefore critically dependent on signals that are contained in the hypervariable domain but can be supported by N-terminal myristoylation or C-terminal prenylation. Interestingly, oncogenic Ras G12V that is localized correctly to the plasma membrane leads to mitogen-activated protein kinase activation irrespective of the combination of targeting signals used for localization, whereas Ras G12V that is mislocalized to the cytosol or to other membranes activates mitogen-activated protein kinase only if the Ras protein is farnesylated.
Databáze: OpenAIRE