Actin isoform utilization during differentiation and remodeling of BC3H1 myogenic cells.
Mouse BC3H1 myogenic cells and a bi-functional chemical cross linking reagent were utilized to investigate the polymerization of newly-synthesized vascular smooth muscle (alpha-actin) and non-muscle (beta- and gamma-actin) actin monomers into native F-actin filament structures during myogenesis. Two actin dimer species were identified by SDS-PAGE analysis of phenylenebismaleimide-cross linked fractions of BC3H1 myoblasts and myocytes. P-dimer was derived from the F-actin-enriched, detergent-insoluble cytoskeleton. Pulse-chase analysis revealed that D-dimer initially was associated with the cytoskeleton but then accumulated in the soluble fraction of lysed muscle cells that contained a non-filamentous or aggregated actin pool. Immunoblot analysis indicated that non-muscle and smooth muscle actins were capable of forming both types of dimer. However, induction of smooth muscle alpha-actin in developing myoblasts coincided with an increase in D-dimer level which may facilitate actin stress fiber assembly. Smooth muscle alpha-actin was rapidly utilized in differentiating myoblasts to assemble extraction-resistant F-actin filaments in the cytoskeleton whereas non-muscle beta- and gamma-actin filaments were more readily dissociated from the cytoskeleton by an extraction buffer containing ATP and EGTA. The data indicate that cytoarchitectural remodeling in developing BC3H1 myogenic cells is accompanied by selective actin isoform utilization that effectively segregates multiple isoactins into different sub-cellular domains and/or supramolecular entities.
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Department of Cell Biology, Neurobiology, and Anatomy, Ohio State University, College of Medicine, Columbus 43210-1239, USA.
Yan
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Strauch
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D000199:Actins / Q000096:biosynthesis / Q000737:chemistry / Q000378:metabolism*; D000818:Animals; D002454:Cell Differentiation; D002478:Cells, Cultured; D003432:Cross-Linking Reagents; D003599:Cytoskeleton / Q000378:metabolism / Q000648:ultrastructure; D019281:Dimerization; D004591:Electrophoresis, Polyacrylamide Gel; D051379:Mice; D009131:Muscle, Smooth, Vascular / Q000378:metabolism; D009132:Muscles / Q000166:cytology* / Q000378:metabolism*; D010449:Peptide Mapping
D000199:Actins; D003432:Cross-Linking Reagents