title: Phosphorylation of the NF2 tumor suppressor in Schwann cells is mediated by Cdc42-Pak and requires paxillin binding.
abstract: Mutations in the Neurofibromatosis type 2 tumor suppressor gene that encodes Schwannomin causes formation of benign schwannomas. Schwannoma cells lose their characteristic bipolar shape and become rounded with excessive ruffling membranes. Schwannomin is phosphorylated at serine 518 (S518) by p21 activated kinase (Pak). Unphosphorylated schwannomin is associated with growth inhibition but little is known about the function of the phosphorylated form, or the molecular events leading to its phosphorylation. Here, we report in SCs that schwannomin S518 phosphorylation requires binding to paxillin and targeting to the plasma membrane. Phospho-S518-schwannomin is enriched in the peripheral-most aspects of membrane specializations where paxillin, activated Pak, Cdc42 but not Rac are highly expressed. Schwannomin and Pak phosphorylation levels are not reduced in response to lowering Rac-GTP levels with NSC23766. Expression of schwannomin S518A/D-GFP variants each distinctively altered Schwann cell shape and polarity. These results are consistent with tight spatial regulation of S518 phosphorylation at the plasma membrane in a paxillin and Cdc42-Pak dependent manner that leads to local reorganization of the SC cytoskeleton.
authors.affiliation: Biomolecular Research Annex, Department of Molecular Biology and Microbiology, University of Central Florida, Orlando, FL 32826, USA.
authors.forename: Courtney
authors.initials: C
authors.lastname: Thaxton
authors.forename: Jorge
authors.initials: J
authors.lastname: Lopera
authors.forename: Marga
authors.initials: M
authors.lastname: Bott
authors.forename: M Elizabeth
authors.initials: ME
authors.lastname: Baldwin
authors.forename: Poonam
authors.initials: P
authors.lastname: Kalidas
authors.forename: Cristina
authors.initials: C
authors.lastname: Fernandez-Valle
mesh_terms: D000818:Animals; D000831:Animals, Newborn; D002478:Cells, Cultured; D004789:Enzyme Activation; D005786:Gene Expression Regulation; D049452:Green Fluorescent Proteins / Q000378:metabolism; D007150:Immunohistochemistry; D047468:Immunoprecipitation; D016296:Mutagenesis; D016518:Neurofibromatosis 2 / Q000378:metabolism*; D025581:Neurofibromin 2 / Q000378:metabolism*; D051419:Paxillin / Q000378:metabolism*; D010766:Phosphorylation; D011485:Protein Binding; D017346:Protein Serine-Threonine Kinases / Q000378:metabolism*; D051381:Rats; D012583:Schwann Cells / Q000378:metabolism*; D014162:Transfection; D020764:cdc42 GTP-Binding Protein / Q000378:metabolism*
chemical_list: D025581:Neurofibromin 2; D051419:Paxillin; D049452:Green Fluorescent Proteins; D017346:Protein Serine-Threonine Kinases; D020764:cdc42 GTP-Binding Protein