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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. - Highlights: • RhoGDI2 is identified as a potential filamin A (FLNA)-binding partner. • Phosphomimetic mutant, RhoGDI2(Tyr153Glu) interacts with FLNA. • RhoGDI2 phosphorylated (Tyr153) by src kinase does not interact with FLNA. • Mutation of Tyr-153more » to Glu of RhoGDI2 does not mimic phosphorylation. • RhoGDI2(Tyr153Glu) provokes an adventitious interaction with FLNA.« less

Authors:
;  [1];  [2]; ;  [1];  [1]
  1. Hematology Division, Department of Medicine, Brigham and Women's Hospital, Harvard Medical School, Boston MA (United States)
  2. Faculty of Veterinary Medicine and Faculty of Biosciences and Pharmacy, University of Leipzig, Leipzig (Germany)
Publication Date:
OSTI Identifier:
22594181
Resource Type:
Journal Article
Journal Name:
Biochemical and Biophysical Research Communications
Additional Journal Information:
Journal Volume: 469; Journal Issue: 3; Other Information: Copyright (c) 2015 Elsevier Science B.V., Amsterdam, The Netherlands, All rights reserved.; Country of input: International Atomic Energy Agency (IAEA); Journal ID: ISSN 0006-291X
Country of Publication:
United States
Language:
English
Subject:
60 APPLIED LIFE SCIENCES; ACTIN; AMINO ACIDS; BLADDER; FILAMENTS; HYBRID SYSTEMS; HYBRIDIZATION; LIGANDS; METASTASES; MUTANTS; MUTATIONS; NEOPLASMS; PHOSPHORYLATION; YEASTS

Citation Formats

Song, Mia, He, Qianjing, Berk, Benjamin-Andreas, Hartwig, John H., Stossel, Thomas P., and Nakamura, Fumihiko. An adventitious interaction of filamin A with RhoGDI2(Tyr153Glu). United States: N. p., 2016. Web. doi:10.1016/J.BBRC.2015.12.044.
Song, Mia, He, Qianjing, Berk, Benjamin-Andreas, Hartwig, John H., Stossel, Thomas P., & Nakamura, Fumihiko. An adventitious interaction of filamin A with RhoGDI2(Tyr153Glu). United States. doi:10.1016/J.BBRC.2015.12.044.
Song, Mia, He, Qianjing, Berk, Benjamin-Andreas, Hartwig, John H., Stossel, Thomas P., and Nakamura, Fumihiko. Fri . "An adventitious interaction of filamin A with RhoGDI2(Tyr153Glu)". United States. doi:10.1016/J.BBRC.2015.12.044.
@article{osti_22594181,
title = {An adventitious interaction of filamin A with RhoGDI2(Tyr153Glu)},
author = {Song, Mia and He, Qianjing and Berk, Benjamin-Andreas and Hartwig, John H. and Stossel, Thomas P. and Nakamura, Fumihiko},
abstractNote = {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. - Highlights: • RhoGDI2 is identified as a potential filamin A (FLNA)-binding partner. • Phosphomimetic mutant, RhoGDI2(Tyr153Glu) interacts with FLNA. • RhoGDI2 phosphorylated (Tyr153) by src kinase does not interact with FLNA. • Mutation of Tyr-153 to Glu of RhoGDI2 does not mimic phosphorylation. • RhoGDI2(Tyr153Glu) provokes an adventitious interaction with FLNA.},
doi = {10.1016/J.BBRC.2015.12.044},
journal = {Biochemical and Biophysical Research Communications},
issn = {0006-291X},
number = 3,
volume = 469,
place = {United States},
year = {2016},
month = {1}
}