A Calcium Signaling Cascade Essential for Myosin Thick Filament Assembly in <i>Xenopus</i> Myocytes

Ferrari, Michael B.; Ribbeck, Katharina; Hagler, Donald J.; Spitzer, Nicholas C.

doi:10.1083/jcb.141.6.1349

Cited by 67 publications

(70 citation statements)

References 59 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…The same cells were relocated and injected with anti-dynein antibody (Sigma clone 70.1) at a needle concentration of 30-50 mg/ml, anti-kinesin HD antibody (gift of Fatima Gyoeva) at a needle concentration of 9.5 mg/ml, anti-HSET antibody at a needle concentration of 10 mg/ml, anti-Eg5 antibody (both gifts of Duane Compton) at a needle concentration of 5.7 mg/ml, nonimmune IgM (Sigma) at a needle concentration of 43 mg/ml, or myosin light chain kinase (MLCK) pseudosubstrate inhibitory peptide (Peptides International) at a needle concentration of 1 mM. The peptide, sequence AKKLSKDRMKKYMARRKWQKTG, specifically inhibits myosin II by acting both as a pseudosubstrate for the autoinhibitory domain of MLCK and by binding to calmodulin and inhibiting Ca 2ϩ /calmodulin-dependent activation of MLCK (12,13).…”

Section: Methodsmentioning

confidence: 99%

Antagonistic forces generated by myosin II and cytoplasmic dynein regulate microtubule turnover, movement, and organization in interphase cells

Yvon

Gross

Wadsworth

2001

Proc. Natl. Acad. Sci. U.S.A.

View full text Add to dashboard Cite

Photoactivation of caged fluorescent tubulin was used mark the microtubule (MT) lattice and monitor MT behavior in interphase cells. A broadening of the photoactivated region occurred as MTs moved bidirectionally. MT movement was not inhibited when MT assembly was suppressed with nocodazole or Taxol; MT movement was suppressed by inhibition of myosin light chain kinase with ML7 or by a peptide inhibitor. Conversely, MT movement was increased after inhibition of cytoplasmic dynein with the antibody 70.1. In addition, the half-time for MT turnover was decreased in cells treated with ML7. These results demonstrate that myosin II and cytoplasmic dynein contribute to a balance of forces that regulates MT organization, movement, and turnover in interphase cells. microtubule motion T he capacity for microtubules (MTs) to undergo dynamic rearrangement is critical to the various vital cellular activities to which they contribute, including the assembly and function of the mitotic spindle, intracellular transport of organelles and vesicles, and the establishment and maintenance of cell shape and polarity. MTs are nucleated at the centrosome and maintain a generally radial organization, with plus-ends at the cell periphery. Direct observations show that peripheral plus-ends undergo dynamic instability behavior, characterized by the stochastic switching between phases of elongation and rapid shortening (1). Dynamic instability is tempered, such that turnover is more rapid in the peripheral region of the cell (2). MT turnover in the more central cytoplasmic domain is thought to occur by subunit turnover at both plus-and minus-ends (3, 4).MTs modulate the behavior of the actin cytoskeleton in a temporally and spatially regulated manner. For example, MTs are required to establish the site of contractile ring formation in cytokinesis and to restrict ruffling behavior to the leading edge of motile fibroblasts (5). The nature of the cross-talk between the MT and actin cytoskeletons is not well understood, but recent work indicates that a feedback mechanism involving Rho family members may contribute to these regulatory interactions (6). In addition, molecular motors may integrate the MT and actin systems. Actomyosin moves MTs rearward at the cell periphery (3), and myosin-generated forces are responsible for MT transport in motile cells and for axon retraction in cultured neurons (7,8). Cytoplasmic dynein contributes to MT organization in interphase cells, to the assembly and function of the mitotic spindle, and to MT transport in neurons (9, 10). These observations led us to test the hypothesis that molecular motors modulate the organization, rearrangement, and turnover of MTs in interphase cells. Materials and MethodsMaterials. All materials for cell culture were obtained from Life Technologies (Gaithersburg, MD), with the exception of FCS, which was obtained from Atlanta Biologicals (Norcross, GA). Unless otherwise noted, all other chemicals were obtained from Sigma.Cell Culture, Cell Staining, and Microinjection. Cell culture...

show abstract

Section: Methodsmentioning

confidence: 99%

Antagonistic forces generated by myosin II and cytoplasmic dynein regulate microtubule turnover, movement, and organization in interphase cells

Yvon

Gross

Wadsworth

2001

Proc. Natl. Acad. Sci. U.S.A.

View full text Add to dashboard Cite

show abstract

“…1E,G). Ryanodine does not reduce cell numbers, disrupt bipolar morphology, alter baseline Ca 2ϩ , or perturb aspects of electrical excitability (Ferrari et al, 1998).…”

Section: Calcium Transients Are Required For Thin Filament Assemblymentioning

confidence: 99%

“…Spontaneous Ca 2ϩ transients are necessary for myosin formation, as demonstrated by blocking their production with bath application of ryanodine (Ferrari et al, 1998). To determine whether transients are required for actin thin filament assembly, we examined actin organization in cells treated with 100 M ryanodine.…”

Section: Calcium Transients Are Required For Thin Filament Assemblymentioning

confidence: 99%

“…On the whole-cell level, it is clear that Ca 2ϩ transients and the resultant contractile activity control major aspects of myofibril assembly, sarcomeric protein turnover, and myofibril maintenance in both cardiac and skeletal myocytes (Sharp et al, 1993;Simpson et al, 1995Simpson et al, , 1996Ferrari et al, 1996Ferrari et al, , 1998Byron et al, 1996;Eble et al, 1998;recent reviews: Berchtold et al, 2000;Gregorio and Antin, 2000;Clark et al, 2002). For example, Ca 2ϩ transients occur spontaneously during embryonic striated muscle development (Flucher and Andrews, 1993;Ferrari et al, 1996;Lorenzon et al, 1997;Ferrari and Spitzer, 1999) and appear to activate MLCK to enable myosin thick filament assembly (Ferrari et al, 1998;Aoki et al, 2000). Whether transients regulate the assembly of other sarcomeric proteins during early myofibrillogenesis has not been examined.…”

Section: Introductionmentioning

confidence: 99%

“…For example, capZ and tropomodulin cap actin filaments at the Z line and pointed end, respectively, and interfering with either capping protein alters normal thin filament assembly (Fowler et al, 1993;Schafer et al, 1993Schafer et al, , 1995Fowler, 1996;Gregorio, 1997;Littlefield and Fowler, 1998). Thick filament assembly, in turn, depends on phosphorylation of the regulatory light chain (RLC) of myosin II by myosin light chain kinase (MLCK; Trybus, 1994;Ferrari and Spitzer, 1998;Aoki et al, 2000). Because thick filaments are organized with a considerable delay compared with the initial periodic organization of I-Z-I complexes LoRusso et al, 1997;Ehler et al, 1999;Van der Ven et al, 1999), interactions between actin and myosin appear unnecessary for the initial establishment of sarcomeric periodicity.…”

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Calcium transients regulate patterned actin assembly during myofibrillogenesis

Cook

Terry

et al. 2003

Developmental Dynamics

Self Cite

View full text Add to dashboard Cite

The highly ordered arrangement of sarcomeric myosin during striated muscle development requires spontaneous calcium (Ca 2ϩ ) transients. Here, we show that blocking transients also compromises patterned assembly of actin thin filaments, titin, and capZ. Because a conserved temporal assembly pattern has been described for these proteins, selective inhibitors of either thick or thin filament formation were used to determine their relative temporal interdependencies.

show abstract

Development of electrical excitability in embryonic neurons: Mechanisms and roles

Spitzer

Ribera

1998

J. Neurobiol.

Self Cite

View full text Add to dashboard Cite

Xenopus spinal neurons serve as a nearly ideal population of excitable cells for study of developmental regulation of electrical excitability. On the one hand, the firing properties of these neurons can be directly examined at early stages of differentiation and membrane excitability changes as neurons mature. Underlying changes in voltage‐dependent ion channels have been characterized and the mechanisms that bring about these changes are being defined. On the other hand, these neurons have been shown to be spontaneously active at stages when action potentials provide significant calcium entry. Calcium entry provokes further elevation of intracellular calcium via release from intracellular stores. The resultant transient elevations of intracellular calcium encode differentiation in their frequency. Recent studies have shown that different neuronal subpopulations enlist distinct mechanisms for regulation of excitability and recruit specific programs of differentiation by particular patterns of activity. © 1998 John Wiley & Sons, Inc. J Neurobiol 37: 190–197, 1998

show abstract

A Calcium Signaling Cascade Essential for Myosin Thick Filament Assembly in Xenopus Myocytes

Cited by 67 publications

References 59 publications

Antagonistic forces generated by myosin II and cytoplasmic dynein regulate microtubule turnover, movement, and organization in interphase cells

Antagonistic forces generated by myosin II and cytoplasmic dynein regulate microtubule turnover, movement, and organization in interphase cells

Calcium transients regulate patterned actin assembly during myofibrillogenesis

Development of electrical excitability in embryonic neurons: Mechanisms and roles

Contact Info

Product

Resources

About