title: Treponema denticola major outer sheath protein induces actin assembly at free barbed ends by a PIP2-dependent uncapping mechanism in fibroblasts.
abstract: The major outer sheath protein (Msp) of Treponema denticola perturbs actin dynamics in fibroblasts by inducing actin reorganization, including subcortical actin filament assembly, leading to defective calcium flux, diminished integrin engagement of collagen, and retarded cell migration. Yet, its mechanisms of action are unknown. We challenged Rat-2 fibroblasts with enriched native Msp. Msp activated the small GTPases Rac1, RhoA and Ras, but not Cdc42, yet only Rac1 localized to areas of actin rearrangement. We used Rac1 dominant negative transfection and chemical inhibition of phosphatidylinositol-3 kinase (PI3K) to show that even though Rac1 activation was PI3K-dependent, neither was required for Msp-induced actin rearrangement. Actin free barbed end formation (FBE) by Msp was also PI3K-independent. Immunoblotting experiments showed that gelsolin and CapZ were released from actin filaments, whereas cofilin remained in an inactive state. Msp induced phosphatidylinositol (4,5)-bisphosphate (PIP2) formation through activation of a phosphoinositide 3-phosphatase and its recruitment to areas of actin assembly at the plasma membrane. Using a PIP2 binding peptide or lipid phosphatase inhibitor, PIP2 was shown to be required for Msp-mediated actin uncapping and FBE formation. Evidently, Msp induces actin assembly in fibroblasts by production and recruitment of PIP2 and release of the capping proteins CapZ and gelsolin from actin barbed ends.
authors: Matrix Dynamics Group, Dental Research Institute, University of Toronto, Toronto, Ontario, Canada. Michelle.visser@utoronto.ca
authors: Michelle B
authors: MB
authors: Visser
authors: Adeline
authors: A
authors: Koh
authors: Michael
authors: M
authors: Glogauer
authors: Richard P
authors: RP
authors: Ellen
mesh_terms: D000199:Actins / Q000378:metabolism*; D000818:Animals; D001426:Bacterial Proteins / Q000494:pharmacology*; D005347:Fibroblasts / Q000187:drug effects* / Q000378:metabolism*; D005455:Fluorescent Antibody Technique; D019269:Phosphatidylinositol 4,5-Diphosphate / Q000378:metabolism*; D018272:Porins / Q000494:pharmacology*; D051381:Rats; D020830:rac1 GTP-Binding Protein / Q000235:genetics / Q000378:metabolism
chemical_list: D000199:Actins; D001426:Bacterial Proteins; D019269:Phosphatidylinositol 4,5-Diphosphate; D018272:Porins; C099361:major outer sheath protein, Treponema; D020830:rac1 GTP-Binding Protein