Plakoglobin is required for maintenance of the cortical actin skeleton in early Xenopus embryos and for cdc42-mediated wound healing.
Early Xenopus embryos are large, and during the egg to gastrula stages, when there is little extracellular matrix, the cytoskeletons of the individual blastomeres are thought to maintain their spherical architecture and provide scaffolding for the cellular movements of gastrulation. We showed previously that depletion of plakoglobin protein during the egg to gastrula stages caused collapse of embryonic architecture. Here, we show that this is due to loss of the cortical actin skeleton after depletion of plakoglobin, whereas the microtubule and cytokeratin skeletons are still present. As a functional assay for the actin skeleton, we show that wound healing, an actin-based behavior in embryos, is also abrogated by plakoglobin depletion. Both wound healing and the amount of cortical actin are enhanced by overexpression of plakoglobin. To begin to identify links between plakoglobin and the cortical actin polymerization machinery, we show here that the Rho family GTPase cdc42, is required for wound healing in the Xenopus blastula. Myc-tagged cdc42 colocalizes with actin in purse-strings surrounding wounds. Overexpression of cdc42 dramatically enhances wound healing, whereas depletion of maternal cdc42 mRNA blocks it. In combinatorial experiments we show that cdc42 cannot rescue the effects of plakoglobin depletion, showing that plakoglobin is required for cdc42-mediated cortical actin assembly during wound healing. However, plakoglobin does rescue the effect of cdc42 depletion, suggesting that cdc42 somehow mediates the distribution or function of plakoglobin. Depletion of alpha-catenin does not remove the cortical actin skeleton, showing that plakoglobin does not mediate its effect by its known linkage through alpha-catenin to the actin skeleton. We conclude that in Xenopus, the actin skeleton is a major determinant of cell shape and overall architecture in the early embryo, and that plakoglobin plays an essential role in the assembly, maintenance, or organization of this cortical actin.
Division of Developmental Biology, Children's Hospital Research Foundation, Medical Center, 3333 Burnet Avenue, Cincinnati, OH 45229, USA.
Matthew
M
Kofron
Janet
J
Heasman
Stephanie A
SA
Lang
Christopher C
CC
Wylie
D000199:Actins / Q000502:physiology*; D000818:Animals; D064199:Cdc20 Proteins; D018797:Cell Cycle Proteins / Q000502:physiology; D048429:Cell Size / Q000502:physiology; D003598:Cytoskeletal Proteins / Q000502:physiology*; D003599:Cytoskeleton / Q000502:physiology*; D051180:Desmoplakins; D029721:Drosophila Proteins; D014945:Wound Healing / Q000502:physiology*; D014982:Xenopus laevis; D051178:alpha Catenin; D020764:cdc42 GTP-Binding Protein / Q000502:physiology*; D051185:gamma Catenin
D000199:Actins; D064199:Cdc20 Proteins; D018797:Cell Cycle Proteins; D003598:Cytoskeletal Proteins; D051180:Desmoplakins; D029721:Drosophila Proteins; D051178:alpha Catenin; C577503:cort protein, Drosophila; D051185:gamma Catenin; D020764:cdc42 GTP-Binding Protein