Optogenetic model reveals cell shape regulation through FAK and Fascin

Journal of Cell Science ◽

10.1242/jcs.258321 ◽

2021 ◽

Author(s):

Jean A. Castillo-Badillo ◽

N. Gautam

Keyword(s):

Focal Adhesion Kinase ◽

Focal Adhesion ◽

Cell Shape ◽

Spherical Shape ◽

Cell Types ◽

Experimental Models ◽

Molecular Processes ◽

Different Cell Types ◽

Shape Changes ◽

In Cells

Cell shape regulation is important but the mechanisms that govern shape are not fully understood, in part due to limited experimental models where cell shape changes and underlying molecular processes can be rapidly and non-invasively monitored in real time. Here, we use an optogenetic tool to activate RhoA in the middle of mononucleated macrophages to induce contraction, resulting in a side with the nucleus that retains its shape and a non-nucleated side which was unable to maintain its shape and collapsed. In cells overexpressing focal adhesion kinase (FAK), the non-nucleated side exhibited a wide flat morphology and was similar in adhesion area to the nucleated side. In cells overexpressing fascin, an actin bundling protein, the non-nucleated side assumed a spherical shape and was similar in height to the nucleated side. This effect of fascin was also observed in fibroblasts even without inducing furrow formation. Based on these results, we conclude that FAK and fascin work together to maintain cell shape by regulating adhesion area and height, respectively, in different cell types.

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Focal Adhesion Kinase Regulation by Oxidative Stress in Different Cell Types

IUBMB Life ◽

10.1080/15216540051081038 ◽

2000 ◽

Vol 50 (4) ◽

pp. 291-299 ◽

Cited By ~ 46

Author(s):

Meriem H. Ben Mahdi ◽

Valérie Andrieu ◽

Catherine Pasquier

Keyword(s):

Oxidative Stress ◽

Focal Adhesion Kinase ◽

Focal Adhesion ◽

Cell Types ◽

Kinase Regulation ◽

Different Cell Types

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Focal Adhesion Kinase Regulation by Oxidative Stress in Different Cell Types

IUBMB Life ◽

10.1080/713803721 ◽

2001 ◽

Vol 50 (4) ◽

pp. 291-299 ◽

Cited By ~ 1

Author(s):

Meriem H. Ben Mahdi ◽

ValÉrie Andrieu ◽

Catherine Pasquier

Keyword(s):

Oxidative Stress ◽

Focal Adhesion Kinase ◽

Focal Adhesion ◽

Cell Types ◽

Kinase Regulation ◽

Different Cell Types

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Differential activation of focal adhesion kinase, Rho and Rac by the ninth and tenth FIII domains of fibronectin

Journal of Cell Science ◽

10.1242/jcs.112.17.2937 ◽

1999 ◽

Vol 112 (17) ◽

pp. 2937-2946

Author(s):

N.A. Hotchin ◽

A.G. Kidd ◽

H. Altroff ◽

H.J. Mardon

Keyword(s):

Growth Factor ◽

Focal Adhesion Kinase ◽

Signaling Pathways ◽

Focal Adhesion ◽

Cell Attachment ◽

Cell Types ◽

Integrin Receptors ◽

3T3 Cells ◽

Swiss 3T3 ◽

Kinase Phosphorylation

Fibronectins are widely expressed extracellular matrix ligands that are essential for many biological processes. Fibronectin-induced signaling pathways are elicited in diverse cell types when specific integrin receptors bind to the ninth and tenth FIII domains, FIII9-10. Integrin-mediated signal transduction involves activation of signaling pathways of the growth factor-dependent Ras-related small GTP-binding proteins Rho and Rac, and phosphorylation of focal adhesion kinase. We have dissected the requirement of FIII9 and FIII10 for Rho and Rac activity and phosphorylation of focal adhesion kinase in BHK fibroblasts and Swiss 3T3 cells. We demonstrate that FIII10 supports cell attachment but does not induce phosphorylation of focal adhesion kinase. In Swiss 3T3 cells, growth factor-independent phosphorylation of focal adhesion kinase and downstream adhesion events are dependent upon the presence of FIII9 in the intact FIII9-10 pair, whereas FIII10-mediated focal adhesion kinase phosphorylation requires a synergistic signal from growth factors. Furthermore, FIII10 is able to elicit cellular responses mediated by Rho, but not Rac, whereas FIII9-10 can elicit both Rho- and Rac-mediated responses. We propose that activation of specific integrin subunits by the FIII10 and FIII9-10 ligands elicits distinct signaling events. This may represent a general molecular mechanism for activation of receptor-specific signaling pathways by a multi-domain ligand.

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Focal adhesion kinase PTK2 autophosphorylation is not required for the activation of sodium–hydrogen exchange by decreased cell volume in the preimplantation mouse embryo

Zygote ◽

10.1017/s0967199419000212 ◽

2019 ◽

Vol 27 (3) ◽

pp. 173-179

Author(s):

Jane C. Fenelon ◽

Baozeng Xu ◽

Jay M. Baltz

Keyword(s):

Focal Adhesion Kinase ◽

Cell Volume ◽

Focal Adhesion ◽

Mouse Embryo ◽

Hydrogen Exchange ◽

Cell Types ◽

Mouse Embryos ◽

Cell Stage ◽

Early Embryos ◽

Early Mouse

SummaryRecovery from decreased cell volume is accomplished by a regulated increase of intracellular osmolarity. The acute response is activation of inorganic ion transport into the cell, the main effector of which is the Na+/H+ exchanger NHE1. NHE1 is rapidly activated by a cell volume decrease in early embryos, but how this occurs is incompletely understood. Elucidating cell volume-regulatory mechanisms in early embryos is important, as it has been shown that their dysregulation results in preimplantation developmental arrest. The kinase JAK2 has a role in volume-mediated NHE1 activation in at least some cells, including 2-cell stage mouse embryos. However, while 2-cell embryos show partial inhibition of NHE1 when JAK2 activity is blocked, NHE1 activation in 1-cell embryos is JAK2-independent, implying a requirement for additional signalling mechanisms. As focal adhesion kinase (FAK aka PTK2) becomes phosphorylated and activated in some cell types in response to decreased cell volume, we sought to determine whether it was involved in NHE1 activation in the early mouse embryo. FAK activity requires initial autophosphorylation of a tyrosine residue, Y397. However, FAK Y397 phosphorylation levels were not increased in either 1- or 2-cell embryos after cell volume was decreased. Furthermore, the selective FAK inhibitor PF-562271 did not affect NHE1 activation at concentrations that essentially eliminated Y397 phosphorylation. Thus, autophosphorylation of FAK Y397 does not appear to be required for NHE1 activation induced by a decrease in cell volume in early mouse embryos.

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JNK Signaling as a Key Modulator of Soft Connective Tissue Physiology, Pathology, and Healing

International Journal of Molecular Sciences ◽

10.3390/ijms21031015 ◽

2020 ◽

Vol 21 (3) ◽

pp. 1015 ◽

Cited By ~ 4

Author(s):

Georgia Nikoloudaki ◽

Sarah Brooks ◽

Alexander P. Peidl ◽

Dylan Tinney ◽

Douglas W. Hamilton

Keyword(s):

Wound Healing ◽

Soft Tissue ◽

Cell Biology ◽

Soft Tissues ◽

Tissue Injury ◽

Cell Types ◽

Cell Phenotype ◽

Cell Behavior ◽

Molecular Processes ◽

Different Cell Types

In healthy individuals, the healing of soft tissues such as skin after pathological insult or post injury follows a relatively predictable and defined series of cell and molecular processes to restore tissue architecture and function(s). Healing progresses through the phases of hemostasis, inflammation, proliferation, remodeling, and concomitant with re-epithelialization restores barrier function. Soft tissue healing is achieved through the spatiotemporal interplay of multiple different cell types including neutrophils, monocytes/macrophages, fibroblasts, endothelial cells/pericytes, and keratinocytes. Expressed in most cell types, c-Jun N-terminal kinases (JNK) are signaling molecules associated with the regulation of several cellular processes involved in soft tissue wound healing and in response to cellular stress. A member of the mitogen-activated protein kinase family (MAPK), JNKs have been implicated in the regulation of inflammatory cell phenotype, as well as fibroblast, stem/progenitor cell, and epithelial cell biology. In this review, we discuss our understanding of JNKs in the regulation of cell behaviors related to tissue injury, pathology, and wound healing of soft tissues. Using models as diverse as Drosophila, mice, rats, as well as human tissues, research is now defining important, but sometimes conflicting roles for JNKs in the regulation of multiple molecular processes in multiple different cell types central to wound healing processes. In this review, we focus specifically on the role of JNKs in the regulation of cell behavior in the healing of skin, cornea, tendon, gingiva, and dental pulp tissues. We conclude that while parallels can be drawn between some JNK activities and the control of cell behavior in healing, the roles of JNK can also be very specific modes of action depending on the tissue and the phase of healing.

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Localization of focal adhesion kinase isoforms in cells of the central nervous system

International Journal of Developmental Neuroscience ◽

10.1016/s0736-5748(02)00126-0 ◽

2003 ◽

Vol 21 (2) ◽

pp. 83-93 ◽

Cited By ~ 17

Author(s):

Andrea Contestabile ◽

Dario Bonanomi ◽

Ferran Burgaya ◽

Jean‐Antoine Girault ◽

Flavia Valtorta

Keyword(s):

Central Nervous System ◽

Nervous System ◽

Focal Adhesion Kinase ◽

Focal Adhesion ◽

The Central Nervous System ◽

In Cells

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Conformational Dynamics of the Focal Adhesion Targeting Domain Control Specific Functions of Focal Adhesion Kinase in Cells

Journal of Biological Chemistry ◽

10.1074/jbc.m114.593632 ◽

2014 ◽

Vol 290 (1) ◽

pp. 478-491 ◽

Cited By ~ 18

Author(s):

Gress Kadaré ◽

Nicolas Gervasi ◽

Karen Brami-Cherrier ◽

Heike Blockus ◽

Said El Messari ◽

...

Keyword(s):

Focal Adhesion Kinase ◽

Focal Adhesion ◽

Conformational Dynamics ◽

In Cells

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Formin-Dependent Adhesions are Required for Invasion by Epithelial Tissues

10.1101/124297 ◽

2017 ◽

Author(s):

Tim B. Fessenden ◽

Yvonne Beckham ◽

Mathew Perez-Neut ◽

Aparajita H. Chourasia ◽

Kay F. Macleod ◽

...

Keyword(s):

Cell Motility ◽

Focal Adhesion ◽

Mdck Cells ◽

Collagen Matrix ◽

Cell Types ◽

Branching Morphogenesis ◽

Mouse Tumor ◽

Collective Invasion ◽

Shape Changes ◽

Tumor Organoids

AbstractDeveloping tissues change shape, and tumors initiate spreading, through collective cell motility. Conserved mechanisms by which tissues initiate motility into their surroundings are not known. We investigated cytoskeletal regulators during collective invasion by mouse tumor organoids and epithelial MDCK acini undergoing branching morphogenesis. Inhibition of formins, but not Arp2/3, prevented the formation of migrating cell fronts in both cell types. MDCK cells depleted of the formin protein Dia1 formed polarized acini and could execute planar cell motility, either within the acinus or in 2D scattering assays. However, Dia1 was required to form protrusions into the collagen matrix. Live imaging of actin, myosin, and collagen in control acini revealed adhesions that deformed individual collagen fibrils, while Dia1-depleted acini exhibited unstable adhesions with minimal collagen deformation. This work identifies Dia1-mediated adhesions as essential regulators of tissue shape changes, through their role in focal adhesion maturation.

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