Organization of stress fibers in cultured fibroblasts after extraction of actin with bovine brain gelsolin-like protein

Verkhovsky, Alexander B.; Surgucheva, Irina; Svitkina, Tatyana; Tint, I.S.; Gelfand, Vladimir I.

doi:10.1016/0014-4827(87)90349-1

Cited by 24 publications

(17 citation statements)

References 11 publications

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“…The following rabbit polyclonal antibodies were used: p16-Arc (Vignjevic et al , 2003), Arp3 (Santa Cruz Biotechnology, Santa Cruz, CA), and p34-Arc subunits of Arp2/3 complex (Millipore, Billerica, MA), nonmuscle myosin II from bovine spleen (Verkhovsky et al , 1987), and capping protein (from D. Schafer, University of Virginia). The following mouse monoclonal antibodies were used: fascin (Millipore), microtubule-associated protein (MAP2) (Sigma-Aldrich, St. Louis, MO), N-cadherin (Santa Cruz Biotechnology; gift from W. J. Nelson, Stanford University, Stanford, CA), α-tubulin (Sigma-Aldrich), and PSD-95 (Abcam, Cambridge, MA).…”

Section: Methodsmentioning

confidence: 99%

Molecular Architecture of Synaptic Actin Cytoskeleton in Hippocampal Neurons Reveals a Mechanism of Dendritic Spine Morphogenesis

Korobova

Svitkina

2010

MBoC

369

381

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Section: Methodsmentioning

confidence: 99%

Molecular Architecture of Synaptic Actin Cytoskeleton in Hippocampal Neurons Reveals a Mechanism of Dendritic Spine Morphogenesis

Korobova

Svitkina

2010

MBoC

369

381

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“…Rat-tail collagen was purchased from BD Biosciences (San Diego, CA). The following primary antibodies were used: mouse anti–cadherin-5 monoclonal antibody (1:200; BD Biosciences), rabbit polyclonal antibody to nonmuscle myosin II from bovine spleen (1:10; Verkhovsky et al , 1987), mouse monoclonal anti-fascin (1:100, clone 55K2; Chemicon, Temecula, CA), rabbit anti-VASP (1:500; gift of Frank Gertler, Massachusetts Institute of Technology, Cambridge, MA), mouse monoclonal anti-tubulin (1:200, clone DM1A; Sigma-Aldrich, St. Louis, MO). Secondary fluorescently labeled antibodies were from Molecular Probes (Invitrogen, Carlsbad, CA) or Jackson ImmunoResearch Laboratories (West Grove, PA).…”

Section: Methodsmentioning

confidence: 99%

The cytoskeletal mechanisms of cell–cell junction formation in endothelial cells

Hoelzle

Svitkina

2012

MBoC

135

120

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“…Fluorescently labeled phalloidin was from Molecular Probes. The following primary antibodies were used: mouse monoclonal α-actinin (Cytoskeleton), mouse monoclonal vinculin (Sigma), rabbit polyclonal NMII from bovine spleen [88], rabbit polyclonal mono- (Ser19) or double- (Thr18/Ser19) phosphorylated MRLC (Cell Signaling Technology, a gift from Dr. C. Chen, University of Pennsylvania). Secondary fluorescently labeled antibodies were from Molecular Probes or Jackson Laboratories.…”

Section: Methodsmentioning

confidence: 99%

Functions of Nonmuscle Myosin II in Assembly of the Cellular Contractile System

et al. 2012

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The contractile system of nonmuscle cells consists of interconnected actomyosin networks and bundles anchored to focal adhesions. The initiation of the contractile system assembly is poorly understood structurally and mechanistically, whereas system’s maturation heavily depends on nonmuscle myosin II (NMII). Using platinum replica electron microscopy in combination with fluorescence microscopy, we characterized the structural mechanisms of the contractile system assembly and roles of NMII at early stages of this process. We show that inhibition of NMII by a specific inhibitor, blebbistatin, in addition to known effects, such as disassembly of stress fibers and mature focal adhesions, also causes transformation of lamellipodia into unattached ruffles, loss of immature focal complexes, loss of cytoskeleton-associated NMII filaments and peripheral accumulation of activated, but unpolymerized NMII. After blebbistatin washout, assembly of the contractile system begins with quick and coordinated recovery of lamellipodia and focal complexes that occurs before reappearance of NMII bipolar filaments. The initial formation of focal complexes and subsequent assembly of NMII filaments preferentially occurred in association with filopodial bundles and concave actin bundles formed by filopodial roots at the lamellipodial base. Over time, accumulating NMII filaments help to transform the precursor structures, focal complexes and associated thin bundles, into stress fibers and mature focal adhesions. However, semi-sarcomeric organization of stress fibers develops at much slower rate. Together, our data suggest that activation of NMII motor activity by light chain phosphorylation occurs at the cell edge and is uncoupled from NMII assembly into bipolar filaments. We propose that activated, but unpolymerized NMII initiates focal complexes, thus providing traction for lamellipodial protrusion. Subsequently, the mechanical resistance of focal complexes activates a load-dependent mechanism of NMII polymerization in association with attached bundles, leading to assembly of stress fibers and maturation of focal adhesions.

show abstract

Organization of stress fibers in cultured fibroblasts after extraction of actin with bovine brain gelsolin-like protein

Cited by 24 publications

References 11 publications

Molecular Architecture of Synaptic Actin Cytoskeleton in Hippocampal Neurons Reveals a Mechanism of Dendritic Spine Morphogenesis

Molecular Architecture of Synaptic Actin Cytoskeleton in Hippocampal Neurons Reveals a Mechanism of Dendritic Spine Morphogenesis

The cytoskeletal mechanisms of cell–cell junction formation in endothelial cells

Functions of Nonmuscle Myosin II in Assembly of the Cellular Contractile System

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