Detection of cellular proteins that interact with the NF2 tumor suppressor gene product.
The neurofibromatosis type 2 (NF2) gene was recently cloned, and the protein it encodes (merlin) was revealed to belong to a family of proteins that link cytoskeletal components with proteins in the cell membrane. To elucidate the biological function of merlin, we produced a bacterial fusion protein consisting of glutathione S-transferase and merlin and used it to detect five merlin-binding cellular proteins, designated p165, p145, p125, p85 and p70, by a protein-binding assay. p165 and merlin were phosphorylated on serine/threonine residues, and immunoprecipitation showed that p85 bound the native form of merlin. Although the entire merlin-ezrin-radixin-moesin (MERM) homology domain of merlin was found to be essential for binding to all five proteins, the MERM homology domains of ezrin and moesin did not bind to any of the five proteins. Since most reported NF2 mutations are in the region we determined was necessary for binding, the mutations probably impair binding. Therefore, the formation of the protein complex is probably crucial for tumor suppression.
Department of Neuro-Onocology, University of Texas M.D. Anderson Cancer Center, Houston 77030.
H
H
Takeshima
I
I
Izawa
P S
PS
Lee
N
N
Safdar
V A
VA
Levin
H
H
Saya
D001483:Base Sequence; D002352:Carrier Proteins / Q000032:analysis* / Q000378:metabolism; D002478:Cells, Cultured; D016515:Genes, Neurofibromatosis 2; D005982:Glutathione Transferase / Q000378:metabolism; D006801:Humans; D008565:Membrane Proteins / Q000032:analysis / Q000378:metabolism*; D008969:Molecular Sequence Data; D009154:Mutation; D009363:Neoplasm Proteins / Q000032:analysis / Q000378:metabolism*; D025581:Neurofibromin 2; D010766:Phosphorylation; D011993:Recombinant Fusion Proteins / Q000378:metabolism
D002352:Carrier Proteins; D008565:Membrane Proteins; D009363:Neoplasm Proteins; D025581:Neurofibromin 2; D011993:Recombinant Fusion Proteins; D005982:Glutathione Transferase