The CaaX motif is required for isoprenylation, carboxyl methylation, and nuclear membrane association of lamin B2.
Recent evidence suggests that the conserved COOH-terminal CaaX motif of nuclear lamins may play a role in targeting newly synthesized proteins to the nuclear envelope. We have shown previously that in rabbit reticulocyte lysates the cysteine residue of the CaaX motif of chicken lamin B2 is necessary for incorporation of a derivative of mevalonic acid, the precursor of isoprenoids. Here we have analyzed the properties of normal and mutated forms of chicken lamin B2 stably expressed in mouse L cells. Mutation of the cysteine residue of the CaaX motif to alanine or introduction of a stop codon immediately after the cysteine residue was found to abolish both isoprenylation and carboxyl methylation of transfected lamin B2. Concomitantly, although nuclear import of the mutant lamin B2 proteins was preserved, their association with the inner nuclear membrane was severely impaired. From these results we conclude that the COOH-terminal CaaX motif is required for isoprenylation and carboxyl methylation of lamins in vivo, and that these modifications are important for association of B-type lamins with the nucleoplasmic surface of the inner nuclear membrane.
Kitten
G T
GT
Swiss Institute for Experimental Cancer Research (ISREC), Epalinges.
Nigg
E A
EA
D000595:Amino Acid Sequence; D000818:Animals; D015153:Blotting, Western; D002451:Cell Compartmentation; D002645:Chickens; D003545:Cysteine; D004252:DNA Mutational Analysis; D005455:Fluorescent Antibody Technique; D066298:In Vitro Techniques; D034921:Lamin Type B; D034882:Lamins; D008745:Methylation; D008798:Mevalonic Acid / Q000378:metabolism*; D008969:Molecular Sequence Data; D009685:Nuclear Envelope / Q000378:metabolism*; D009687:Nuclear Proteins / Q000737:chemistry / Q000378:metabolism*; D011106:Polyisoprenyl Phosphates / Q000378:metabolism; D011499:Protein Processing, Post-Translational; D011994:Recombinant Proteins / Q000378:metabolism; D012995:Solubility
D034921:Lamin Type B; D034882:Lamins; D009687:Nuclear Proteins; D011106:Polyisoprenyl Phosphates; D011994:Recombinant Proteins; D003545:Cysteine; D008798:Mevalonic Acid