Regulation of intracellular actin polymerization by prenylated cellular proteins.
Posttranslational modification by covalent attachment of polyisoprene intermediates to a carboxyterminal CAAX-box motif is required for the biologic function of proteins such as p21ras, the supergene family of ras-related proteins, nuclear lamins, and subunits of heterotrimeric G-proteins. Cells grown in the presence of lovastatin, which inhibits HMG-CoA reductase and prevents synthesis of intermediates required for protein prenylation, develop a round, refractile morphology. Our data indicate that this is due to the selective loss of actin cables without gross changes in the microtubular lattice or intermediate filament structure. Microinjection of a competitive peptide inhibitor of protein prenyltransferases into the cytoplasm of cells induces an identical change in morphology with loss of actin cables. Mevalonate (MVA) reverses the lovastatin-induced morphologic change by inducing a rapid repolymerization of actin cables with coincident reversion to the flat morphology. Furthermore, microinjection of farnesyl-pyrophosphate or geranylgeranyl-pyrophosphate into lovastatin-treated cells also results in rapid morphologic reversion. The morphologic reversion induced by MVA requires the presence of serum, and is independent of extracellular calcium. The addition of cycloheximide to the growth medium prevents lovastatin-induced loss of actin cables, and causes morphologic reversion of lovastatin-treated cells by a mechanism that is independent of MVA. A1F4- induces morphologic reversion in a manner indistinguishable from MVA. These data indicate that prenylated protein(s) play a critical role in regulating the state of intracellular actin, and that GGPP can rescue the lovastatin-induced morphologic phenotype in the absence of upstream intermediates of cholesterol biosynthesis. We have begun to dissect the signaling events that mediate this pathway.
Fenton
R G
RG
NCI-FCRDC, Biological Response Modifiers Program, Frederick, Maryland 21702.
Kung
H F
HF
Longo
D L
DL
Smith
M R
MR
D008841:Actin Cytoskeleton / Q000187:drug effects / Q000648:ultrastructure*; D000199:Actins / Q000378:metabolism*; D001769:Blood; D002118:Calcium / Q000494:pharmacology; D002460:Cell Line; D004122:Dimethylallyltranstransferase / Q000378:metabolism; D006801:Humans; D008148:Lovastatin / Q000494:pharmacology*; D008798:Mevalonic Acid / Q000494:pharmacology; D011106:Polyisoprenyl Phosphates / Q000494:pharmacology; D011108:Polymers; D011499:Protein Processing, Post-Translational; D011506:Proteins / Q000378:metabolism*; D012717:Sesquiterpenes
D000199:Actins; D011106:Polyisoprenyl Phosphates; D011108:Polymers; D011506:Proteins; D012717:Sesquiterpenes; C004808:farnesyl pyrophosphate; D008148:Lovastatin; D004122:Dimethylallyltranstransferase; C002963:geranylgeranyl pyrophosphate; D008798:Mevalonic Acid; D002118:Calcium