Association of the growth-arrest-specific protein Gas7 with F-actin induces reorganization of microfilaments and promotes membrane outgrowth.
The growth-arrest-specific gene, Gas7, is required for neurite outgrowth in cerebellar neurons. Here we report that Gas7 can induce the formation of extended cellular processes in NIH3T3 cells by interacting with actin and mediating reorganization of microfilaments. The Gas 7 protein, which increased markedly during growth arrest of NIH3T3 cells and persisted transiently at high levels upon reentry of cells into the cell cycle, localized near the plasma membrane and selectively colocalized with microfilaments in membrane ruffles. Process extensions induced by ectopic overexpression of Gas7 were blocked by the actin-depolymerizing agent cytochalasin D, suggesting that membrane extensions produced by Gas7 require actin polymerization. Association of endogenous Gas7 protein with microfilaments was verified by F-actin affinity chromatography; direct binding of purified His-Gas7 to actin also was demonstrated and shown to be mediated by the Gas7 C-terminal domain. Similarly, localization of Gas7 in membrane ruffles was mediated by the C-terminal domain, although neither this region nor the N-terminal domain was individually sufficient to induce process formation. Biochemical studies and electron microscopy showed that both full-length Gas7 protein and its C-terminal region can promote actin assembly as well as the crosslinking of actin filaments. We propose that Gas7 localized near the plasma membrane induces the assembly of actin and the membrane outgrowth.
She
Bin-Ru
BR
Institute of Molecular Biology, Academia Sinica, Nankang, Taipei, 115, Taiwan.
Liou
Gunn-Guang
GG
Lin-Chao
Sue
S
D016475:3T3 Cells / Q000378:metabolism; D008841:Actin Cytoskeleton / Q000187:drug effects / Q000378:metabolism* / Q000648:ultrastructure; D000199:Actins / Q000378:metabolism*; D000818:Animals; D001665:Binding Sites; D015152:Blotting, Northern; D001940:Breast / Q000502:physiology; D002453:Cell Cycle; D002462:Cell Membrane / Q000502:physiology; D002645:Chickens; D002846:Chromatography, Affinity; D015638:Cytochalasin D / Q000494:pharmacology; D003599:Cytoskeleton / Q000187:drug effects / Q000378:metabolism; D005434:Flow Cytometry; D015151:Immunoblotting; D051379:Mice; D008854:Microscopy, Electron; D008856:Microscopy, Fluorescence; D009419:Nerve Tissue Proteins / Q000235:genetics / Q000378:metabolism*
D000199:Actins; C485435:Gas7 protein, mouse; D009419:Nerve Tissue Proteins; D015638:Cytochalasin D