title: An adventitious interaction of filamin A with RhoGDI2(Tyr153Glu).
abstract: Filamin A (FLNA) is an actin filament crosslinking protein with multiple intracellular binding partners. Mechanical force exposes cryptic FLNA binding sites for some of these ligands. To identify new force-dependent binding interactions, we used a fusion construct composed of two FLNA domains, one of which was previously identified as containing a force-dependent binding site as a bait in a yeast two-hybrid system and identified the Rho dissociation inhibitor 2 (RhoGDI2) as a potential interacting partner. A RhoGDI2 truncate with 81 N-terminal amino acid residues and a phosphomimetic mutant, RhoGDI(Tyr153Glu) interacted with the FLNA construct. However, neither wild-type or full-length RhoGDI2 phosphorylated at Y153 interacted with FLNA. Our interpretation of these contradictions is that truncation and/or mutation of RhoGDI2 perturbs its conformation to expose a site that adventitiously binds FLNA and is not a bona-fide interaction. Therefore, previous studies reporting that a RhoGDI(Y153E) mutant suppresses the metastasis of human bladder cancer cells must be reinvestigated in light of artificial interaction of this point mutant with FLNA.
authors.affiliation: Hematology Division, Department of Medicine, Brigham and Women's Hospital, Harvard Medical School, Boston MA, USA.
authors.forename: Mia
authors.initials: M
authors.lastname: Song
authors.affiliation: Hematology Division, Department of Medicine, Brigham and Women's Hospital, Harvard Medical School, Boston MA, USA.
authors.forename: Qianjing
authors.initials: Q
authors.lastname: He
authors.affiliation: Faculty of Veterinary Medicine and Faculty of Biosciences and Pharmacy, University of Leipzig, Leipzig, Germany.
authors.forename: Benjamin-Andreas
authors.initials: BA
authors.lastname: Berk
authors.affiliation: Hematology Division, Department of Medicine, Brigham and Women's Hospital, Harvard Medical School, Boston MA, USA.
authors.forename: John H
authors.initials: JH
authors.lastname: Hartwig
authors.affiliation: Hematology Division, Department of Medicine, Brigham and Women's Hospital, Harvard Medical School, Boston MA, USA.
authors.forename: Thomas P
authors.initials: TP
authors.lastname: Stossel
authors.affiliation: Hematology Division, Department of Medicine, Brigham and Women's Hospital, Harvard Medical School, Boston MA, USA. Electronic address: fnakamura@partners.org.
authors.forename: Fumihiko
authors.initials: F
authors.lastname: Nakamura
mesh_terms: D001665:Binding Sites; D064448:Filamins / Q000737:chemistry* / Q000378:metabolism*; D057809:HEK293 Cells; D006801:Humans; D010766:Phosphorylation; D011485:Protein Binding; D062949:rho Guanine Nucleotide Dissociation Inhibitor beta / Q000737:chemistry* / Q000378:metabolism*
chemical_list: C569658:ARHGDIB protein, human; C578663:FLNA protein, human; D064448:Filamins; D062949:rho Guanine Nucleotide Dissociation Inhibitor beta
keywords: Filamin; Mechanotransduction; Phosphomimetic mutant; Phosphorylation; RhoGDI2; Src