Myosin II filament assemblies in the active lamella of fibroblasts: their morphogenesis and role in the formation of actin filament bundles.
The morphogenesis of myosin II structures in active lamella undergoing net protrusion was analyzed by correlative fluorescence and electron microscopy. In rat embryo fibroblasts (REF 52) microinjected with tetramethylrhodamine-myosin II, nascent myosin spots formed close to the active edge during periods of retraction and then elongated into wavy ribbons of uniform width. The spots and ribbons initially behaved as distinct structural entities but subsequently aligned with each other in a sarcomeric-like pattern. Electron microscopy established that the spots and ribbons consisted of bipolar minifilaments associated with each other at their head-containing ends and arranged in a single row in an "open" zig-zag conformation or as a "closed" parallel stack. Ribbons also contacted each other in a nonsarcomeric, network-like arrangement as described previously (Verkhovsky and Borisy, 1993. J. Cell Biol. 123:637-652). Myosin ribbons were particularly pronounced in REF 52 cells, but small ribbons and networks were found also in a range of other mammalian cells. At the edge of the cell, individual spots and open ribbons were associated with relatively disordered actin filaments. Further from the edge, myosin filament alignment increased in parallel with the development of actin bundles. In actin bundles, the actin cross-linking protein, alpha-actinin, was excluded from sites of myosin localization but concentrated in paired sites flanking each myosin ribbon, suggesting that myosin filament association may initiate a pathway for the formation of actin filament bundles. We propose that zig-zag assemblies of myosin II filaments induce the formation of actin bundles by pulling on an actin filament network and that co-alignment of actin and myosin filaments proceeds via folding of myosin II filament assemblies in an accordion-like fashion.
D016475:3T3 Cells / Q000737:chemistry / Q000502:physiology / Q000648:ultrastructure; D000185:Actinin / Q000032:analysis; D000199:Actins / Q000032:analysis / Q000096:biosynthesis* / Q000648:ultrastructure; D000818:Animals; D002460:Cell Line / Q000737:chemistry / Q000502:physiology / Q000648:ultrastructure; D002461:Cell Line, Transformed / Q000502:physiology; D002642:Chick Embryo; D002522:Chlorocebus aethiops; D005347:Fibroblasts / Q000737:chemistry / Q000502:physiology* / Q000648:ultrastructure; D005455:Fluorescent Antibody Technique; D006801:Humans; D007150:Immunohistochemistry; D051379:Mice; D016253:Microscopy, Immunoelectron; D009024:Morphogenesis / Q000502:physiology; D009218:Myosins / Q000502:physiology* / Q000648:ultrastructure; D051381:Rats; D013997:Time Factors; D014422:Turkeys
D000199:Actins; D000185:Actinin; D009218:Myosins