14-3-3 Coordinates Microtubules, Rac, and Myosin II to Control Cell Mechanics and Cytokinesis

Zhou, Qiongqiong; Kee, Yee Seir; Poirier, Christopher C.; Jelinek, Christine; Osborne, Jonathan; Divi, Srikanth; Surcel, Alexandra; Will, Marie E.; Eggert, Ulrike; Müller‐Taubenberger, Annette; Iglesias, Pablo A.; Cotter, Robert J.; Robinson, Douglas N.

doi:10.1016/j.cub.2010.09.048

Cited by 72 publications

(124 citation statements)

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“…To investigate the role of the assembly domain of myosin in 4-HAP activation further, we performed in vitro assembly assays on a myosin II tail fragment, assembly domain C-terminal, which is sufficient to reconstitute regulatable myosin II BTF assembly, as well as on tail fragments from MYH9 and MYH10. These experiments were also conducted in the presence or absence of 14-3-3, a myosin II-binding partner that sequesters free myosin monomers, thus increasing the sensitivity of the assembly assay and providing a positive control for a direct effector of myosin II assembly (8). In all experimental setups, 4-HAP did not affect the assembly of myosin II, including the MYH9 and MYH10 paralogs (Fig.…”

Section: Resultsmentioning

confidence: 99%

“…2D). Because myosin II is a known effector of cell mechanics, both in Dictyostelium as well as in other organisms (8,(10)(11)(12)(13)(14), we next queried whether the increase in cortical localization would have an impact on the mechanical properties of the cell. Using micropipette aspiration (MPA) assays, we determined that acute treatment with 700 pM carbamate-7 led to a 1.4-fold increase in the cell's cortical tension ( Fig.…”

Section: Resultsmentioning

confidence: 99%

“…To assess whether carbamate-7 affects known cytokinesis pathways, we targeted two spatially distinct modules, one at the equatorial plane of a dividing cell regulated by spindle signals and the mechanosensory system of Myosin II/Cortexillin I, and one at the polar cortex regulated by the RacE/14-3-3/MyoII pathway (8). In chemical-genetic epistasis analyses, we challenged mutant cell lines targeting both modules with carbamate-7.…”

Section: Resultsmentioning

confidence: 99%

“…For example, 95% of PDACs have early activating mutations in Kras, which modulates cell elasticity (23,24). Early PanINs also up-regulate the actin cross-linking protein fascin, whereas later stages are marked by the up-regulation of 14-3-3σ, a regulator of myosin II assembly (8,25,26). Furthermore, serial analysis of gene expression of numerous Pancs that were compared with normal pancreatic cells (HPDE cells) revealed alterations in the expression of several regulators of myosin II assembly and contractility (9).…”

Section: Small Molecule 4-hap Stiffens Pancreatic Cancer Cells and Hementioning

confidence: 99%

“…Carbamate-7 is a highly potent cell mechanics modulator that inhibits cytokinesis weakly, as designed by our specific experimental approach. Evaluation of carbamate-7 demonstrated that it inhibits cytokinesis by influencing the RacE/14-3-3/Myosin II (MyoII) pathway, known to regulate cortical mechanics (8). This activity arises not from carbamate-7, but rather from the action of two degradation products: 3,4-dichloroaniline (3,4-DCA) and 4-hydroxyacetophenone (4-HAP).…”

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Pharmacological activation of myosin II paralogs to correct cell mechanics defects

Surcel¹,

Ng²,

West‐Foyle³

et al. 2015

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

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Current approaches to cancer treatment focus on targeting signal transduction pathways. Here, we develop an alternative system for targeting cell mechanics for the discovery of novel therapeutics. We designed a live-cell, high-throughput chemical screen to identify mechanical modulators. We characterized 4-hydroxyacetophenone (4-HAP), which enhances the cortical localization of the mechanoenzyme myosin II, independent of myosin heavy-chain phosphorylation, thus increasing cellular cortical tension. To shift cell mechanics, 4-HAP requires myosin II, including its full power stroke, specifically activating human myosin IIB (MYH10) and human myosin IIC (MYH14), but not human myosin IIA (MYH9). We further demonstrated that invasive pancreatic cancer cells are more deformable than normal pancreatic ductal epithelial cells, a mechanical profile that was partially corrected with 4-HAP, which also decreased the invasion and migration of these cancer cells. Overall, 4-HAP modifies nonmuscle myosin II-based cell mechanics across phylogeny and disease states and provides proof of concept that cell mechanics offer a rich drug target space, allowing for possible corrective modulation of tumor cell behavior. mechanical modulator | 3,4-dichloroaniline | 4-hydroxyacetophenone | myosin II | pancreatic cancer C ell shape change processes include cell growth, division, motility, and the formation of complex structures like tissues and organs, all of which are governed by the intersection of biochemistry, genetics, and mechanics. These three modules are integral not just for normal function in healthy cells but also in disease states. Pharmacological manipulation of some of these modules has already led to treatment strategies for inflammation and cancer (e.g., paclitaxel) (1, 2). However, many presently available therapies, which address only one aspect of cell shape change, typically either fail to abolish the disease completely or lead to compensatory regulatory changes, and therefore to drug resistance. Targeting cell mechanics remains an underused approach for drug development. In cancer, altered cell mechanics are a hallmark of metastatic efficiency: cell stiffness decreases up to 70% in many metastatic cancer cells (3-5). It is rational then that one therapeutic approach is to increase cellular elasticity, which would, in turn, reduce metastatic potential and act downstream of cancer-inducing genetic alterations.Known mechanical modulators (e.g., latrunculin, blebbistatin) are often lethal, have numerous off-site targets, and act to generate a softer and metastatic-like mechanical phenotype (6, 7). However, the field's ability to increase cellular elasticity on acute time scales is highly restricted. In an effort to close this gap and find modulators that stiffen cells, we leveraged our molecular and analytical understanding of cytokinesis, an evolutionarily conserved and highly mechanical cell shape change event, to establish an in vivo, large-scale, high-throughput chemical screen for small-molecule modulators of cell shap...

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Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Section: Small Molecule 4-hap Stiffens Pancreatic Cancer Cells and Hementioning

confidence: 99%

mentioning

confidence: 99%

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Pharmacological activation of myosin II paralogs to correct cell mechanics defects

Surcel¹,

Ng²,

West‐Foyle³

et al. 2015

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

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Microtubule Organisation inDictyostelium

Gräf

2015

Encyclopedia of Life Sciences

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Dictyostelium amoebae contain a radial array of microtubules emanating from a single microtubule‐organising centre called centrosome that is bound to the cytosolic face of the nucleus. Their centrosome contains no centrioles but consists of a layered core surrounded by a corona harbouring microtubule nucleation centres. It duplicates in prophase of a closed mitosis and organises a central spindle that drives centrosome separation and chromosome segregation. Dictyostelium microtubules exhibit a differential dynamic behaviour during interphase. Growth and shrinkage is observed only in the periphery but not in the pericentrosomal region. During mitosis, when centrosomes possess no corona, microtubules behave quite dynamically in formation of a central spindle and astral microtubules. Microtubules are associated with a couple of conserved proteins ( MAPs ), which are involved in centrosome biogenesis and the cross talk of microtubule tips with the actin cell cortex. The latter becomes evident in cytokinesis, when centrosomes with their attached microtubules participate in the positioning of cleavage furrows. Key Concepts Dictyostelium amoebae contain a nucleus‐associated centrosome that serves as the only microtubule‐organising centre. The Dictyostelium centrosome contains no centrioles, but consists of a three‐layered core structure surrounded by a microtubule‐nucleating corona. If compared to the three major plaques of the yeast spindle pole body, the entire core structure of the Dictyostelium centrosome appears equivalent to the central plaque, while the corona plays a similar role as the inner and outer plaques. Dictyostelium centrosomes duplicate at the onset of mitosis. Dictyostelium amoebae show a closed type of mitosis with a persisting nuclear envelope. Dictyostelium microtubules are quite dynamic during mitosis but show growth and shrinkage only in the periphery during interphase. Microtubule plus ends regulate actin dynamics at the cell cortex. Dictyostelium amoebae are a useful model to study the role of the centrosome and microtubules in cell dynamics and disease.

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Cytokinesis mechanics and mechanosensing

West‐Foyle

Robinson

2012

Cytoskeleton

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Cytokinesis shape change occurs through the interfacing of three modules, cell mechanics, myosin II-mediated contractile stress generation and sensing, and a control system of regulatory proteins, which together ensure flexibility and robustness. This integrated system then defines the stereotypical shape changes of successful cytokinesis, which occurs under a diversity of mechanical contexts and environmental conditions.

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14-3-3 Coordinates Microtubules, Rac, and Myosin II to Control Cell Mechanics and Cytokinesis

Cited by 72 publications

References 34 publications

Pharmacological activation of myosin II paralogs to correct cell mechanics defects

Pharmacological activation of myosin II paralogs to correct cell mechanics defects

Microtubule Organisation inDictyostelium

Cytokinesis mechanics and mechanosensing

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