Evidence for a cytoskeleton attachment domain at the N-terminus of the NF2protein

Neurofibromatosis type 2 is a hereditary cancer syndrome characterized by t he development of bilateral vestibular schwannomas. Underlying the disease are inactivating mutations of the NF2 tumor suppressor gene, located on chr omosome 22, encoding a 595-amino-acid protein. The NF2 protein, also known as merlin or schwannomin, is reported to act as a membrane-cytoskeleton lin king protein. This assumption is based on the homology of the NF2 protein t o a group of band 4.1-related proteins, ezrin, radixin, and moesin. The cyt oskeletal association of the NF2 protein has in part been confirmed by its ability to resist extraction from cells by nonionic detergents. We performe d detergent extraction on COS cells transfected with NF2 cDNA constructs. T he extracts were analyzed by Western blotting and immunofluorescent stainin g with monoclonal anti-NF2 antibodies. The results provide evidence for a h igh-affinity cytoskeleton attachment domain at amino acids 29-131 and a put ative lower affinity domain between amino acids 321 and 470. (C) 2000 Wiley -Liss, Inc.