Nonmuscle myosin IIA and IIB differently suppress microtubule growth to stabilize cell morphology

2019 ◽  
Vol 167 (1) ◽  
pp. 25-39 ◽  
Author(s):  
Yuta Sato ◽  
Keiju Kamijo ◽  
Motosuke Tsutsumi ◽  
Yota Murakami ◽  
Masayuki Takahashi
Keyword(s):  
Cell Morphology ◽  
Control Cell ◽  
Growth Dynamics ◽  
Actin Dynamics ◽  
Nonmuscle Myosin ◽  
Cellular Processes ◽  
Nonmuscle Myosin Ii ◽  
Cytoskeletal Dynamics ◽  
Dependent Inhibition ◽  
The Stability

Abstract Precise regulation of cytoskeletal dynamics is important in many fundamental cellular processes such as cell shape determination. Actin and microtubule (MT) cytoskeletons mutually regulate their stability and dynamics. Nonmuscle myosin II (NMII) is a candidate protein that mediates the actin–MT crosstalk. NMII regulates the stability and dynamics of actin filaments to control cell morphology. Additionally, previous reports suggest that NMII-dependent cellular contractility regulates MT dynamics, and MTs also control cell morphology; however, the detailed mechanism whereby NMII regulates MT dynamics and the relationship among actin dynamics, MT dynamics and cell morphology remain unclear. The present study explores the roles of two well-characterized NMII isoforms, NMIIA and NMIIB, on the regulation of MT growth dynamics and cell morphology. We performed RNAi and drug experiments and demonstrated the NMII isoform-specific mechanisms—NMIIA-dependent cellular contractility upregulates the expression of some mammalian diaphanous-related formin (mDia) proteins that suppress MT dynamics; NMIIB-dependent inhibition of actin depolymerization suppresses MT growth independently of cellular contractility. The depletion of either NMIIA or NMIIB resulted in the increase in cellular morphological dynamicity, which was alleviated by the perturbation of MT dynamics. Thus, the NMII-dependent control of cell morphology significantly relies on MT dynamics.

scholarly journals Overlapping Roles of Drosophila Drak and Rok Kinases in Epithelial Tissue Morphogenesis

2010 ◽  
Vol 21 (16) ◽  
pp. 2869-2879 ◽  
Author(s):  
Dagmar Neubueser ◽  
David R. Hipfner
Keyword(s):  
Rho Gtpase ◽  
Actin Dynamics ◽  
Signaling Networks ◽  
Epithelial Tissue ◽  
Nonmuscle Myosin ◽  
Dynamic Regulation ◽  
Epithelial Tissues ◽  
Nonmuscle Myosin Ii ◽  
Death Associated Protein Kinase

Dynamic regulation of cytoskeletal contractility through phosphorylation of the nonmuscle Myosin-II regulatory light chain (MRLC) provides an essential source of tension for shaping epithelial tissues. Rho GTPase and its effector kinase ROCK have been implicated in regulating MRLC phosphorylation in vivo, but evidence suggests that other mechanisms must be involved. Here, we report the identification of a single Drosophila homologue of the Death-associated protein kinase (DAPK) family, called Drak, as a regulator of MRLC phosphorylation. Based on analysis of null mutants, we find that Drak broadly promotes proper morphogenesis of epithelial tissues during development. Drak activity is largely redundant with that of the Drosophila ROCK orthologue, Rok, such that it is essential only when Rok levels are reduced. We demonstrate that these two kinases synergistically promote phosphorylation of Spaghetti squash (Sqh), the Drosophila MRLC orthologue, in vivo. The lethality of drak/rok mutants can be rescued by restoring Sqh activity, indicating that Sqh is the critical common effector of these two kinases. These results provide the first evidence that DAPK family kinases regulate actin dynamics in vivo and identify Drak as a novel component of the signaling networks that shape epithelial tissues.

scholarly journals Distinct roles of nonmuscle myosin II isoforms for establishing tension and elasticity during cell morphodynamics

2020 ◽  
Author(s):  
Kai Weißenbruch ◽  
Justin Grewe ◽  
Kathrin Stricker ◽  
Laurent Baulesch ◽  
Ulrich S. Schwarz ◽  
...  
Keyword(s):  
Mammalian Cells ◽  
Myosin Ii ◽  
Force Generation ◽  
Nonmuscle Myosin ◽  
Cellular Processes ◽  
Scale Bridging ◽  
Crossbridge Cycle ◽  
Nonmuscle Myosin Ii ◽  
Actin Fiber ◽  
And Migration

AbstractNonmuscle myosin II (NM II) is an integral part of essential cellular processes, including adhesion and migration. Mammalian cells express up to three isoforms termed NM IIA, B, and C. We used U2OS cells to create CRISPR/Cas9-based knockouts of all three isoforms and analyzed the phenotypes on homogeneous and micropatterned substrates. We find that NM IIA is essential to build up cellular tension during initial stages of force generation, while NM IIB is necessary to elastically stabilize NM IIA-generated tension. The knockout of NM IIC has no detectable effects. A scale-bridging mathematical model explains our observations by relating actin fiber stability to the molecular rates of the myosin crossbridge cycle. We also find that NM IIA initiates and guides co-assembly of NM IIB into heterotypic minifilaments. We finally use mathematical modeling to explain the different exchange dynamics of NM IIA and B in minifilaments, as measured in FRAP experiments.

scholarly journals Targeting an anchored phosphatase-deacetylase unit restores renal ciliary homeostasis

eLife ◽  
2021 ◽  
Vol 10 ◽  
Author(s):  
Janani Gopalan ◽  
Mitchell H Omar ◽  
Ankita Roy ◽  
Nelly M Cruz ◽  
Jerome Falcone ◽  
...  
Keyword(s):  
Primary Cilia ◽  
Genetic Disorders ◽  
Chronic Kidney Failure ◽  
Actin Dynamics ◽  
Histone Deacetylase 6 ◽  
Collecting Ducts ◽  
Cytoskeletal Dynamics ◽  
Anchoring Protein ◽  
On Chip ◽  
The Stability

Pathophysiological defects in water homeostasis can lead to renal failure. Likewise, common genetic disorders associated with abnormal cytoskeletal dynamics in the kidney collecting ducts and perturbed calcium and cAMP signaling in the ciliary compartment contribute to chronic kidney failure. We show that collecting ducts in mice lacking the A-Kinase anchoring protein AKAP220 exhibit enhanced development of primary cilia. Mechanistic studies reveal that AKAP220-associated protein phosphatase 1 (PP1) mediates this phenotype by promoting changes in the stability of histone deacetylase 6 (HDAC6) with concomitant defects in actin dynamics. This proceeds through a previously unrecognized adaptor function for PP1 as all ciliogenesis and cytoskeletal phenotypes are recapitulated in mIMCD3 knock-in cells expressing a phosphatase-targeting defective AKAP220-ΔPP1 mutant. Pharmacological blocking of local HDAC6 activity alters cilia development and reduces cystogenesis in kidney-on-chip and organoid models. These findings identify the AKAP220-PPI-HDAC6 pathway as a key effector in primary cilia development.

scholarly journals Methamphetamine learning induces persistent nonmuscle myosin II-dependent spine motility in the basolateral amygdala

2019 ◽  
Author(s):  
Erica J. Young ◽  
Hua Lin ◽  
Theodore M. Kamenecka ◽  
Gavin Rumbaugh ◽  
Courtney A. Miller
Keyword(s):  
Basolateral Amygdala ◽  
Molecular Mechanisms ◽  
Myosin Ii ◽  
Selective Vulnerability ◽  
Actin Dynamics ◽  
Nonmuscle Myosin ◽  
Systemic Injection ◽  
Nonmuscle Myosin Ii ◽  
Window Of Vulnerability ◽  
Induced Changes

ABSTRACTNonmuscle myosin II inhibition (NMIIi) in the basolateral amygdala (BLA) selectively disrupts memories associated with methamphetamine (METH) days after learning, without retrieval. However, the molecular mechanisms underlying this selective vulnerability remain poorly understood. A known function of NMII is to transiently activate dendritic spine actin dynamics with learning. Therefore, we hypothesized that METH-associated learning perpetuates NMII-driven actin dynamics in dendritic spines, leading to an extended window of vulnerability for memory disruption. Two-photon imaging of actin-mediated spine motility in neurons from memory-related structures, BLA and CA1, revealed a persistent increase in spine motility after METH-associated learning that was restricted to BLA neurons. METH-induced changes to BLA spine dynamics were reversed by a single systemic injection of an NMII inhibitor. Thus, a perpetual form of NMII-driven spine actin dynamics in BLA neurons may contribute to the unique susceptibility of METH-associated memories.

Nonmuscle Myosin II Regulation Directs Its Multiple Roles in Cell Migration and Division

2021 ◽  
Vol 37 (1) ◽  
Author(s):  
Marina Garrido-Casado ◽  
Gloria Asensio-Juárez ◽  
Miguel Vicente-Manzanares
Keyword(s):  
Cell Migration ◽  
Conformational Changes ◽  
Myosin Ii ◽  
Annual Review ◽  
Publication Date ◽  
Dependent Manner ◽  
Control Force ◽  
Nonmuscle Myosin ◽  
Nonmuscle Myosin Ii ◽  
The Stability

Nonmuscle myosin II (NMII) is a multimeric protein complex that generates most mechanical force in eukaryotic cells. NMII function is controlled at three main levels. The first level includes events that trigger conformational changes that extend the complex to enable its assembly into filaments. The second level controls the ATPase activity of the complex and its binding to microfilaments in extended NMII filaments. The third level includes events that modulate the stability and contractility of the filaments. They all work in concert to finely control force generation inside cells. NMII is a common endpoint of mechanochemical signaling pathways that control cellular responses to physical and chemical extracellular cues. Specific phosphorylations modulate NMII activation in a context-dependent manner. A few kinases control these phosphorylations in a spatially, temporally, and lineage-restricted fashion, enabling functional adaptability to the cellular microenvironment. Here, we review mechanisms that control NMII activity in the context of cell migration and division. Expected final online publication date for the Annual Review of Cell and Developmental Biology, Volume 37 is October 2021. Please see http://www.annualreviews.org/page/journal/pubdates for revised estimates.

scholarly journals RhoD regulates cytoskeletal dynamics via the actin nucleation–promoting factor WASp homologue associated with actin Golgi membranes and microtubules

2012 ◽  
Vol 23 (24) ◽  
pp. 4807-4819 ◽  
Author(s):  
Annica K. B. Gad ◽  
Vishal Nehru ◽  
Aino Ruusala ◽  
Pontus Aspenström
Keyword(s):  
Cell Migration ◽  
Actin Filament ◽  
Rho Gtpases ◽  
Actin Dynamics ◽  
Filamin A ◽  
Actin Nucleation ◽  
Cellular Processes ◽  
Golgi Membranes ◽  
Cytoskeletal Dynamics ◽  
Nucleation Promoting Factor

The Rho GTPases have mainly been studied in association with their roles in the regulation of actin filament organization. These studies have shown that the Rho GTPases are essential for basic cellular processes, such as cell migration, contraction, and division. In this paper, we report that RhoD has a role in the organization of actin dynamics that is distinct from the roles of the better-studied Rho members Cdc42, RhoA, and Rac1. We found that RhoD binds the actin nucleation–promoting factor WASp homologue associated with actin Golgi membranes and microtubules (WHAMM), as well as the related filamin A–binding protein FILIP1. Of these two RhoD-binding proteins, WHAMM was found to bind to the Arp2/3 complex, while FILIP1 bound filamin A. WHAMM was found to act downstream of RhoD in regulating cytoskeletal dynamics. In addition, cells treated with small interfering RNAs for RhoD and WHAMM showed increased cell attachment and decreased cell migration. These major effects on cytoskeletal dynamics indicate that RhoD and its effectors control vital cytoskeleton-driven cellular processes. In agreement with this notion, our data suggest that RhoD coordinates Arp2/3-dependent and FLNa-dependent mechanisms to control the actin filament system, cell adhesion, and cell migration.

scholarly journals Targeting an anchored phosphatase-deacetylase unit restores renal ciliary homeostasis

2021 ◽  
Author(s):  
Janani Gopalan ◽  
Mitch Omar ◽  
Ankita Roy ◽  
Nelly M. Cruz ◽  
Jerome Falcone ◽  
...  
Keyword(s):  
Primary Cilia ◽  
Genetic Disorder ◽  
Actin Dynamics ◽  
Histone Deacetylase 6 ◽  
Collecting Ducts ◽  
Cytoskeletal Dynamics ◽  
Anchoring Protein ◽  
Autosomal Dominant Polycystic Kidney ◽  
On Chip ◽  
The Stability

AbstractPathophysiological defects in water homeostasis can lead to renal failure. Autosomal dominant polycystic kidney disease (ADPKD) is a common genetic disorder associated with abnormal cytoskeletal dynamics in the kidney collecting ducts and perturbed calcium and cAMP signaling in the ciliary compartment. We show that collecting ducts in mice lacking the A-Kinase anchoring protein AKAP220 exhibit enhanced development of primary cilia. Mechanistic studies reveal that AKAP220-associated protein phosphatase 1 (PP1) mediates this phenotype by promoting changes in the stability of histone deacetylase 6 (HDAC6) with concomitant defects in actin dynamics. This proceeds through a previously unrecognized adaptor function for PP1 as all ciliogenesis and cytoskeletal phenotypes are recapitulated in mIMCD3 knock-in cells expressing a phosphatase-targeting defective AKAP220-ΔPP1 mutant. Pharmacological blocking of local HDAC6 activity alters cilia development and reduces cystogenesis in kidney-on-chip and organoid models of ADPKD. These findings identify the AKAP220-PPI-HDAC6 pathway as a key effector in primary cilia development.

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