Post-translational processing of RhoA. Carboxyl methylation of the carboxyl-terminal prenylcysteine increases the half-life of Rhoa.
RhoA and related GTP-binding proteins are modified post-translationally at their carboxyl terminus to form a prenylcysteine methyl ester. The synthesis and post-translational modification of RhoA and Cdc42 were examined in the RAW264 macrophage cell line, and the effect of carboxyl methylation on protein turnover was determined. Cells were labeled with [35S]cysteine, and RhoA or Cdc42 was immunoprecipitated with specific antibodies. Both RhoA and Cdc42 were methylated rapidly in control cells, with little accumulation of unmethylated protein. Carboxyl methylation of RhoA was inhibited by incubation of cells with a carbocyclic adenosine analog, 3-deazaaristeromycin, resulting in the accumulation of unmethylated RhoA. Under these conditions, Cdc42 methylation was inhibited only partially. When methylation was inhibited, the RhoA half-life decreased from 31 to 12 h, and the Cdc42 half-life decreased from 15 to 11 h. The increased degradation of unmethylated RhoA demonstrates a novel function for carboxyl-terminal prenylcysteine carboxyl methylation in protecting RhoA and related proteins from degradation.
Backlund
P S
PS
Laboratory of General and Comparative Biochemistry, National Institute of Mental Health, Bethesda, Maryland 20892, USA. backlund@helix.nih.gov
D000818:Animals; D018797:Cell Cycle Proteins / Q000378:metabolism; D002460:Cell Line; D003545:Cysteine / Q000031:analogs & derivatives* / Q000378:metabolism; D019204:GTP-Binding Proteins / Q000378:metabolism*; D006207:Half-Life; D007700:Kinetics; D008264:Macrophages / Q000378:metabolism; D008745:Methylation; D051379:Mice; D011499:Protein Processing, Post-Translational; D020764:cdc42 GTP-Binding Protein; D020742:rhoA GTP-Binding Protein
D018797:Cell Cycle Proteins; D019204:GTP-Binding Proteins; D020764:cdc42 GTP-Binding Protein; D020742:rhoA GTP-Binding Protein; D003545:Cysteine