title: GEF-H1 controls focal adhesion signaling that regulates mesenchymal stem cell lineage commitment.
abstract: Focal adhesions (FAs) undergo maturation that culminates in size and composition changes that modulate adhesion, cytoskeleton remodeling and differentiation. Although it is well recognized that stimuli for osteogenesis of mesenchymal stem cells (MSCs) drive FA maturation, actin organization and stress fiber polarization, the extent to which FA-mediated signals regulated by the FA protein composition specifies MSC commitment remains largely unknown. Here, we demonstrate that, upon dexamethasone (osteogenic induction) treatment, guanine nucleotide exchange factor H1 (GEF-H1, also known as Rho guanine nucleotide exchange factor 2, encoded by ARHGEF2) is significantly enriched in FAs. Perturbation of GEF-H1 inhibits FA formation, anisotropic stress fiber orientation and MSC osteogenesis in an actomyosin-contractility-independent manner. To determine the role of GEF-H1 in MSC osteogenesis, we explore the GEF-H1-modulated FA proteome that reveals non-muscle myosin-II heavy chain-B (NMIIB, also known as myosin-10, encoded by MYH10) as a target of GEF-H1 in FAs. Inhibition of targeting NMIIB into FAs suppresses FA formation, stress fiber polarization, cell stiffness and osteogenic commitments in MSCs. Our data demonstrate a role for FA signaling in specifying MSC commitment.
authors.affiliation: Institute of Biochemistry and Molecular Biology, National Yang-Ming University, Taipei 11221, Taiwan.
authors.forename: I-Husan
authors.initials: IH
authors.lastname: Huang
authors.affiliation: Institute of Biochemical Sciences, National Taiwan University, Taipei 10617, Taiwan Institute of Biological Chemistry, Academia Sinica, Taipei 11529, Taiwan.
authors.forename: Cheng-Te
authors.initials: CT
authors.lastname: Hsiao
authors.affiliation: Institute of Biochemistry and Molecular Biology, National Yang-Ming University, Taipei 11221, Taiwan.
authors.forename: Jui-Chung
authors.initials: JC
authors.lastname: Wu
authors.affiliation: Proteomics and Analytical Biochemistry Unit, Research Resources Branch, National Institute on Aging, NIH, Baltimore, MD 21224, USA jckuo@ym.edu.tw.
authors.forename: Rong-Fong
authors.initials: RF
authors.lastname: Shen
authors.affiliation: Institute of Basic Medical Sciences, National Cheng Kung University, Tainan 70101, Taiwan Department of Physiology, National Cheng Kung University, Tainan 70101, Taiwan.
authors.forename: Ching-Yi
authors.initials: CY
authors.lastname: Liu
authors.affiliation: Department of Cell Biology and Anatomy, National Cheng Kung University, Tainan 70101, Taiwan Center for Neurotrauma and Neuroregeneration, Taipei Medical University, Taipei 11031, Taiwan.
authors.forename: Yang-Kao
authors.initials: YK
authors.lastname: Wang
authors.affiliation: Institute of Biochemistry and Molecular Biology, National Yang-Ming University, Taipei 11221, Taiwan.
authors.forename: Yu-Chen
authors.initials: YC
authors.lastname: Chen
authors.affiliation: Institute of Biochemistry and Molecular Biology, National Yang-Ming University, Taipei 11221, Taiwan.
authors.forename: Chi-Ming
authors.initials: CM
authors.lastname: Huang
authors.affiliation: Institute of Engineering in Medicine, University of California at San Diego, La Jolla, CA 92093, USA Department of Mechanical and Aerospace Engineering, University of California at San Diego, La Jolla, CA 92093, USA.
authors.forename: Juan C
authors.initials: JC
authors.lastname: del Álamo
authors.affiliation: Institute of Biochemistry and Molecular Biology, National Yang-Ming University, Taipei 11221, Taiwan.
authors.forename: Zee-Fen
authors.initials: ZF
authors.lastname: Chang
authors.affiliation: Institute of Basic Medical Sciences, National Cheng Kung University, Tainan 70101, Taiwan Department of Physiology, National Cheng Kung University, Tainan 70101, Taiwan.
authors.forename: Ming-Jer
authors.initials: MJ
authors.lastname: Tang
authors.affiliation: Institute of Biochemical Sciences, National Taiwan University, Taipei 10617, Taiwan Institute of Biological Chemistry, Academia Sinica, Taipei 11529, Taiwan.
authors.forename: Kay-Hooi
authors.initials: KH
authors.lastname: Khoo
authors.affiliation: Institute of Biochemistry and Molecular Biology, National Yang-Ming University, Taipei 11221, Taiwan jckuo@ym.edu.tw.
authors.forename: Jean-Cheng
authors.initials: JC
authors.lastname: Kuo
mesh_terms: D002454:Cell Differentiation / Q000502:physiology; D019070:Cell Lineage; D022001:Focal Adhesions / Q000378:metabolism*; D006801:Humans; D059630:Mesenchymal Stem Cells / Q000166:cytology* / Q000378:metabolism*; D010012:Osteogenesis; D064067:Rho Guanine Nucleotide Exchange Factors / Q000378:metabolism*; D015398:Signal Transduction
chemical_list: D064067:Rho Guanine Nucleotide Exchange Factors
keywords: Focal adhesions; Mesenchymal stem cell; Osteogenesis; Stress fiber
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