Dynamin2 organizes lamellipodial actin networks to orchestrate lamellar actomyosin.
Actin networks in migrating cells exist as several interdependent structures: sheet-like networks of branched actin filaments in lamellipodia; arrays of bundled actin filaments co-assembled with myosin II in lamellae; and actin filaments that engage focal adhesions. How these dynamic networks are integrated and coordinated to maintain a coherent actin cytoskeleton in migrating cells is not known. We show that the large GTPase dynamin2 is enriched in the distal lamellipod where it regulates lamellipodial actin networks as they form and flow in U2-OS cells. Within lamellipodia, dynamin2 regulated the spatiotemporal distributions of α-actinin and cortactin, two actin-binding proteins that specify actin network architecture. Dynamin2's action on lamellipodial F-actin influenced the formation and retrograde flow of lamellar actomyosin via direct and indirect interactions with actin filaments and a finely tuned GTP hydrolysis activity. Expression in dynamin2-depleted cells of a mutant dynamin2 protein that restores endocytic activity, but not activities that remodel actin filaments, demonstrated that actin filament remodeling by dynamin2 did not depend of its functions in endocytosis. Thus, dynamin2 acts within lamellipodia to organize actin filaments and regulate assembly and flow of lamellar actomyosin. We hypothesize that through its actions on lamellipodial F-actin, dynamin2 generates F-actin structures that give rise to lamellar actomyosin and for efficient coupling of F-actin at focal adhesions. In this way, dynamin2 orchestrates the global actin cytoskeleton.
Department of Biology, University of Virginia, Charlottesville, Virginia, United States of America.
Manisha
M
Menon
Department of Biology, University of Virginia, Charlottesville, Virginia, United States of America.
Olga L
OL
Askinazi
Department of Biology, University of Virginia, Charlottesville, Virginia, United States of America; Department of Cell Biology, University of Virginia, Charlottesville, Virginia, United States of America.
Dorothy A
DA
Schafer
D008841:Actin Cytoskeleton / Q000378:metabolism; D000199:Actins / Q000378:metabolism*; D000205:Actomyosin / Q000378:metabolism*; D000818:Animals; D002448:Cell Adhesion; D002460:Cell Line; D034285:Dynamin II / Q000737:chemistry / Q000378:metabolism*; D006160:Guanosine Triphosphate / Q000378:metabolism; D006868:Hydrolysis; D017434:Protein Structure, Tertiary; D021381:Protein Transport; D011554:Pseudopodia / Q000378:metabolism*; D051381:Rats
D000199:Actins; D006160:Guanosine Triphosphate; D000205:Actomyosin; D034285:Dynamin II