title: Establishment of stable human fibroblast cell lines constitutively expressing active Rho-GTPases.
abstract: Small GTP-binding proteins of the Rho family (RhoA, Cdc42, Rac1) regulate the organisation and the turnover of the cell's cytoskeleton and adhesion structures. A significant function of these cellular structures is to translate and counterbalance forces applied to, or generated by, cells in order to maintain homeostasis and control cell movement. We therefore hypothesised that Rho-GTPases are directly involved in cellular gravity perception and may participate in the alterations induced in microgravity. To define an adequate cellular model allowing to investigate this issue, we have established stable cell lines constitutively expressing active forms of either RhoA, Cdc42, or Rac1. The three cell lines differ by morphology and by their ability to form filopodia, lamellipodia, and bundles of actin stress fibers. Overexpression of the active form of either RhoA, Cdc42, or Rac1 is compatible with cell viability and does not affect cell population doubling time. Thus, our series of mutant cells appear well suited to gain further knowledge on the molecular mechanisms of cellular gravity perception.
authors.affiliation: Laboratory of Connective Tissues Biology, Centre de Recherche de la Groupement Interdisciplinaire de Génomique Appliquée, University of Liège, Liège, Belgium.
authors.forename: S
authors.initials: S
authors.lastname: Servotte
authors.forename: Z
authors.initials: Z
authors.lastname: Zhang
authors.forename: C A
authors.initials: CA
authors.lastname: Lambert
authors.forename: T T Giang
authors.initials: TT
authors.lastname: Ho
authors.forename: G
authors.initials: G
authors.lastname: Chometon
authors.forename: B
authors.initials: B
authors.lastname: Eckes
authors.forename: T
authors.initials: T
authors.lastname: Krieg
authors.forename: C M
authors.initials: CM
authors.lastname: Lapière
authors.forename: B V
authors.initials: BV
authors.lastname: Nusgens
authors.forename: M
authors.initials: M
authors.lastname: Aumailley
mesh_terms: D008841:Actin Cytoskeleton / Q000378:metabolism; D002460:Cell Line; D049109:Cell Proliferation; D048430:Cell Shape; D004789:Enzyme Activation; D005347:Fibroblasts / Q000166:cytology / Q000201:enzymology*; D006801:Humans; D009154:Mutation; D011554:Pseudopodia / Q000378:metabolism; D013997:Time Factors; D014162:Transfection; D016596:Vinculin / Q000378:metabolism; D020764:cdc42 GTP-Binding Protein / Q000378:metabolism; D020830:rac1 GTP-Binding Protein / Q000378:metabolism; D020741:rho GTP-Binding Proteins / Q000235:genetics / Q000378:metabolism*; D020742:rhoA GTP-Binding Protein / Q000378:metabolism
chemical_list: C488955:VCL protein, human; D016596:Vinculin; D020764:cdc42 GTP-Binding Protein; D020830:rac1 GTP-Binding Protein; D020741:rho GTP-Binding Proteins; D020742:rhoA GTP-Binding Protein