Abstract
Neurofibromatosis type 2 (NF2) protein, also known as merlin or schwannomin, is a tumor suppressor, and NF2 is mutated in most schwannomas and meningiomas. Although these tumors are dependent on NF2, some lack detectable NF2 mutations, which indicates that alternative mechanisms exist for inactivating merlin. Here, we demonstrate cleavage of merlin by the ubiquitous protease calpain and considerable activation of the calpain system resulting in the loss of merlin expression in these tumors. Increased proteolysis of merlin by calpain in some schwannomas and meningiomas exemplifies tumorigenesis linked to the calpain-mediated proteolytic pathway.
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References
Evans, D.G. et al. A genetic study of type 2 neurofibromatosis in the United Kingdom. J. Med. Genet. 29, 841–846 (1992).
Martuza, R.L., & Eldridge, R. Neurofibromatosis 2 (bilateral acoustic neurofibromatosis). N. Engl. J. Med. 318, 684–688 (1988).
Eldridge, R. Central neurofibromatosis with bilateral acoustic neurinoma. Adv. Neurol. 29, 57–65 (1981).
Rouleau, C.A. et al. Alteration in a new gene encoding a putative membrane-organizing protein causes neurofibromatosis type 2. Nature 363, 515–521 (1993).
Trofatter, J.A. et al. A novel moesin-, ezrin-, radixin-like gene is a candidate for the neurofibromatosis 2 tumor suppressor. Cell 72, 791–800 (1993).
Tsukita, S. & Yonemura, S. ERM (ezrin/radixin/moesin) family: from cytoskeleton to signal transduction. Curr. Opin. Cell Biol. 9, 70–75 (1997).
Algrain, M., Turunen, O., Vaheri, A., Louvard, D. & Arpin, M. Ezrin contains cytoskeleton and membrane binding domains accounting for its proposed role as a membrane-cytoskeletal linker. J. Cell Biol. 120, 129–139 (1993).
Koga, H. et al. Impairment of cell adhesion by expression of the mutant neurofibromatosis type 2(NF2) genes which miss exons in the ERM-homology domain. Oncogene (in the press).
Ruttledge, M.H. et al. Evidence for the complete inactivation of the NF2 gene in the majority of sporadic meningiomas. Nature Genet. 6, 180–184 (1994).
Merel, P. et al. Screening for germ-line mutations in the NF2 gene. Genes Chromosom. Cancer 12, 117–127 (1995).
Lekanne, D.R. et al. Frequent NF2 gene transcript mutations in sporadic meningiomas and vestibular schwannomas. Am. J. Hum. Genet. 54, 1022–1029 (1994).
Harada, T. et al. Molecular genetic investigation of the neurofibromatosis type 2 tumor suppressor gene in sporadic meningioma. J. Neurosurg. 84, 847–851 (1996).
Bijlsma, E.K., Brouwer, M.R., Bosch, D.A., Westerveld, A. & Hulsebos, T.J. Molecular characterization of chromosome 22 deletions in schwannomas. Genes Chromosom. Cancer 5, 201–205 (1992).
Twist, E.C. et al. The neurofibromatosis type 2 gene is inactivated in schwannomas. Hum. Mol. Genet. 3, 147–151 (1994).
Arakawa, H., Hayashi, N., Nagase, H., Ogawa, M. & Nakamura, Y. Alternative splicing of the NF2 gene and its mutation analysis of breast and colorectal cancers. Hum. Mol. Genet. 3, 565–568 (1994).
Bianchi, A.B. et al. High frequency of inactivating mutations in the neurofibromatosis type 2 gene (NF2) in primary malignant mesotheliomas. Proc. Natl. Acad. Sci. USA 92, 10854–10858 (1995).
Sekido, Y. et al. Neurofibromatosis type 2 (NF2) gene is somatically mutated in mesothelioma but not in lung cancer. Cancer Res. 55, 1227–1231 (1995).
MacCollin, M. et al. Mutational analysis of patients with neurofibromatosis 2. Am. J. Hum. Genet. 55, 314–320 (1994).
Welling, D.B. et al. Mutational spectrum in the neurofibromatosis type 2 gene in sporadic and familial schwannomas. Hum. Genet. 98, 189–193 (1996).
Stemmer-Rachamimov, O.A. et al. Universal absence of merlin, but not other ERM family members, in schwannomas. Am. J. Pathol. 151, 1649–1654 (1997).
Gutmann, D.H., Giordano, M.J., Fishback, A.S. & Guha, A. Loss of merlin expression in sporadic meningiomas, ependymomas and schwannomas. Neurology 49, 267–270 (1997).
Lee, J.H. et al. Reduced expression of schwannomin/merlin in human sporadic meningiomas. Neurosurgery 40, 578–587 (1997).
Takeshima, H. et al. Detection of cellular proteins that interact with the NF2 tumor suppressor gene product. Oncogene 9, 2135–2144 (1994).
Saido, T.C., Sorimachi, H. & Suzuki, K. Calpain: new perspectives in molecular diversity and physiological-pathological involvement. FASEB J. 8, 814–822 (1994).
Suzuki, K., Sorimachi, H., Yoshizawa, T., Kinbara, K. & Ishiura, S. Calpain: novel family members, activation, and physiologic function. Biol. Chem. Hoppe-Seyler 376, 523–529 (1995).
Sorimachi, H., Ishiura, S. & Suzuki, K. Structure and physiological function of calpains. Biochem. J. 328, 721–732 (1997).
Tsubuki, S., Saito, Y., Tomioka, M., Ito, H. & Kawashima, S. Differential inhibition of calpain and proteasome activities by peptidyl aldehydes of dileucine and trileucine. J. Biochem. 119, 572–576 (1996).
Coux, O., Tanaka, K. & Goldberg, A.L. Structure and functions of the 20S and 26S proteasomes. Annu. Rev. Biochem. 65, 801–847 (1996).
Burkhart, W.A., Moyer, M.B., Bailey, J.M. & Miller, C.G. Electroblotting of proteins to Teflon tape and membranes for N- and C-terminal sequence analysis. Anal. Biochem. 236, 364–367 (1996).
Miller, C.G. et. al. Techniques in Protein Chemistry Vol. VI, (ed. Crabb, John W.) 219–227 (Academic, San Diego, California, 1995).
Nagao, S. et al. Calpain-calpastatin interactions in epidermoid carcinoma KB cells. J. Biochem. 115, 1178–1184 (1994)
Billger, M., Wallin, M. & Karlsson, J.O. Proteolysis of tubulin and microtubule-associated proteins 1 and 2 by calpain I and II. Difference in sensitivity of assembled and disassembled microtubules. Cell Calcium 9, 33–44 (1988).
Saido, T.C. et al. Autolytic transition of μ-calpain upon activation as resolved by antibodies distinguishing between the pre- and post-autolysis forms. J. Biochem. 111, 81–86 (1992).
Siman, R., Baudry, M. & Lynch, G. Brain fodrin: substrate for calpain I, an endogenous calcium-activated protease. Proc. Natl. Acad. Sci. USA 81, 3572–3576 (1984).
Yang, L.S. & Ksiezak, R.H. Calpain-induced proteolysis of normal human tau and tau associated with paired helical filaments. Eur. J. Biochem. 233, 9–17 (1995).
Inomata, M. et al. Involvement of calpain in integrin-mediated signal transduction. Arch. Biochem. Biophys. 328, 129–134 (1996).
Selliah, N., Brooks, W.H. & Roszman, T.L. Proteolytic cleavage of alpha-actinin by calpain in T cells stimulated with anti-CD3 monoclonal antibody. J. Immunol. 156, 3215–3221 (1996).
Jay, D. & Stracher, A. Expression in Escherichia coli, phosphorylation with cAMP-dependent protein kinase and proteolysis by calpain of a 71-kDa domain of human endothelial actin binding protein. Biochem. Biophys. Res. Commun. 232, 555–558 (1997).
Huang, C. et al. Proteolysis of platelet cortactin by calpain. J. Biol. Chem. 272, 19248–19252 (1997).
Du, X. et al. Calpain cleavage of the cytoplasmic domain of the integrin beta 3 sub-unit. J. Biol. Chem. 270, 26146–26151 (1995).
Elvira, M., Diez, J.A., Wang, K.K. & Villalobo, A. Phosphorylation of connexin-32 by protein kinase C prevents its proteolysis by μ-calpain and m-calpain. J. Biol. Chem. 268, 14294–14300 (1993).
Litersky, J.M. & Johnson, G.V. Phosphorylation by cAMP-dependent protein kinase inhibits the degradation of tau by calpain. J. Biol. Chem. 267, 1563–1568 (1992).
Greenwood, J.A., Troncoso, J.C., Costello, A.C. & Johnson, G.V. Phosphorylation modulates calpain-mediated proteolysis and calmodulin binding of the 200-kDa and 160-kDa neurofilament proteins. J. Neurochem. 61, 191–199 (1993).
Scheffner, M., Huibregtse, J.M., Vierstra, R.D. & Howley, P.M. The HPV-16 E6 and E6-AP complex functions as a ubiquitin-protein ligase in the ubiquitination of p53. Cell 75, 495–505 (1993).
Catzavelos, C. et al. Decreased levels of the cell-cycle inhibitor p27Kpl protein: prognostic implications in primary breast cancer. Nature Med. 3, 227–230 (1997).
Loda, M. et al. Increased proteasome-dependent degradation of the cyclin-dependent kinase inhibitor p27 in aggressive colorectal carcinomas. Nature Med. 3, 231–234 (1997).
Jensen, R.L., Origitano, T.C., Lee, Y.S., Weber, M. & Wurster, R.D. In vitro growth inhibition of growth factor-stimulated meningioma cells by calcium channel antagonists. Neurosurgery 36, 365–373 (1995).
Jensen, R.L. et al. Inhibition of in vitro meningioma proliferation after growth factor stimulation by calcium channel antagonists: Part II-Additional growth factors, growth factor receptor immunohistochemistry, and intracellular calcium measurements. Neurosurgery 37, 937–946 (1995).
Felgner, P.L. et al. Lipofection: a highly efficient, lipid-mediated DNA-transfection procedure. Proc. Natl. Acad. Sci. USA 84, 7413–7417 (1987).
Siebert, P.D. & Chenchik, A. Modified acid guanidinium thiocyanate-phenol-chloroform RNA extraction method which greatly reduces DNA contamination. Nucleic Acids Res. 21, 2019–2020 (1993).
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Kimura, Y., Koga, H., Araki, N. et al. The involvement of calpain-independent proteolysis of the tumor suppressor NF2 (merlin) in schwannomas and meningiomas. Nat Med 4, 915–922 (1998). https://doi.org/10.1038/nm0898-915
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DOI: https://doi.org/10.1038/nm0898-915