Galpha12/13 is essential for directed cell migration and localized Rho-Dia1 function.
Scratch-wound assays are frequently used to study directed cell migration, a process critical for embryogenesis, invasion, and tissue repair. The function and identity of trimeric G-proteins in cell behavior during wound healing is not known. Here we show that Galpha12/13, but not Galphaq/11 or Galphai, is indispensable for coordinated and directed cell migration. In mouse embryonic fibroblasts endogenous Rho activity is present at the rear of migrating cells but also at the leading edge, whereas it is undetectable at the cell front of Galpha12/13-deficient mouse embryonic fibroblasts. Spatial activation of Rho at the wound edge can be stimulated by lysophosphatidic acid. Active Rho colocalizes with the diaphanous-related formin Dia1 at the cell front. Galpha12/13-deficient cells lack Dia1 localization to the wound edge and are unable to form orientated, stable microtubules during wound healing. Knock down of Dia1 reveals its requirement for microtubule stabilization as well as polarized cell migration. Thus, we identified Galpha12/13-proteins as essential components linking extracellular signals to localized Rho-Dia1 function during directed cell movement.
Goulimari
Polyxeni
P
Institute of Pharmacology, University of Heidelberg, Im Neuenheimer Feld 366, 69120 Heidelberg, Germany.
Kitzing
Thomas M
TM
Knieling
Helga
H
Brandt
Dominique T
DT
Offermanns
Stefan
S
Grosse
Robert
R
D000345:Affinity Labels; D000818:Animals; D002352:Carrier Proteins / Q000502:physiology*; D002460:Cell Line; D002465:Cell Movement / Q000502:physiology*; D003599:Cytoskeleton / Q000502:physiology; D005455:Fluorescent Antibody Technique; D000080865:Formins; D044365:GTP-Binding Protein alpha Subunits, G12-G13 / Q000502:physiology*; D051379:Mice; D008870:Microtubules / Q000502:physiology; D034622:RNA Interference; D011994:Recombinant Proteins / Q000378:metabolism; D020741:rho GTP-Binding Proteins / Q000502:physiology*
D000345:Affinity Labels; D002352:Carrier Proteins; C491464:Diap1 protein, mouse; D000080865:Formins; D011994:Recombinant Proteins; D044365:GTP-Binding Protein alpha Subunits, G12-G13; D020741:rho GTP-Binding Proteins