scholarly journals Filamin A is required for vimentin-mediated cell adhesion and spreading

2010 ◽  
Vol 298 (2) ◽  
pp. C221-C236 ◽  
Author(s):  
Hugh Kim ◽  
Fumihiko Nakamura ◽  
Wilson Lee ◽  
Yulia Shifrin ◽  
Pamela Arora ◽  
...  
Keyword(s):  
Protein Kinase C ◽  
Cell Adhesion ◽  
Protein Kinase ◽  
Cell Spreading ◽  
Intermediate Filament Protein ◽  
Β1 Integrin ◽  
Filamin A ◽  
Integrin Activation ◽  
Hek 293 ◽  
Kinase C

Cell adhesion and spreading are regulated by complex interactions involving the cytoskeleton and extracellular matrix proteins. We examined the interaction of the intermediate filament protein vimentin with the actin cross-linking protein filamin A in regulation of spreading in HEK-293 and 3T3 cells. Filamin A and vimentin-expressing cells were well spread on collagen and exhibited numerous cell extensions enriched with filamin A and vimentin. By contrast, cells treated with small interfering RNA (siRNA) to knock down filamin A or vimentin were poorly spread; both of these cell populations exhibited >50% reductions of cell adhesion, cell surface β1 integrin expression, and β1 integrin activation. Knockdown of filamin A reduced vimentin phosphorylation and blocked recruitment of vimentin to cell extensions, whereas knockdown of filamin and/or vimentin inhibited the formation of cell extensions. Reduced vimentin phosphorylation, cell spreading, and β1 integrin surface expression, and activation were phenocopied in cells treated with the protein kinase C inhibitor bisindolylmaleimide; cell spreading was also reduced by siRNA knockdown of protein kinase C-ε. By immunoprecipitation of cell lysates and by pull-down assays using purified proteins, we found an association between filamin A and vimentin. Filamin A also associated with protein kinase C-ε, which was enriched in cell extensions. These data indicate that filamin A associates with vimentin and to protein kinase C-ε, thereby enabling vimentin phosphorylation, which is important for β1 integrin activation and cell spreading on collagen.

scholarly journals Tetraspanin CD82 drives acute myeloid leukemia chemoresistance by modulating protein kinase C alpha and β1 integrin activation

Oncogene ◽  
2020 ◽  
Vol 39 (19) ◽  
pp. 3910-3925
Author(s):  
Muskan Floren ◽  
Sebastian Restrepo Cruz ◽  
Christina M. Termini ◽  
Kristopher D. Marjon ◽  
Keith A. Lidke ◽  
...  
Keyword(s):  
Acute Myeloid Leukemia ◽  
Protein Kinase C ◽  
Protein Kinase ◽  
Myeloid Leukemia ◽  
Β1 Integrin ◽  
Integrin Activation ◽  
Kinase C ◽  
Protein Kinase C Alpha ◽  
Acute Myeloid

Regulation of cell adhesion to collagen via β1 integrins is dependent on interactions of filamin A with vimentin and protein kinase C epsilon

2010 ◽  
Vol 316 (11) ◽  
pp. 1829-1844 ◽  
Author(s):  
Hugh Kim ◽  
Fumihiko Nakamura ◽  
Wilson Lee ◽  
Claire Hong ◽  
Dolores Pérez-Sala ◽  
...  
Keyword(s):  
Protein Kinase C ◽  
Cell Adhesion ◽  
Protein Kinase ◽  
Filamin A ◽  
Kinase C ◽  
Protein Kinase C Epsilon ◽  
Β1 Integrins

scholarly journals Protein kinase C regulates the recruitment of syndecan-4 into focal contacts.

1995 ◽  
Vol 6 (11) ◽  
pp. 1503-1513 ◽  
Author(s):  
P C Baciu ◽  
P F Goetinck
Keyword(s):  
Protein Kinase C ◽  
Cell Adhesion ◽  
Protein Kinase ◽  
Heparan Sulfate ◽  
Heparan Sulfate Proteoglycans ◽  
Matrix Deposition ◽  
Kinase C ◽  
Focal Contacts ◽  
The Matrix ◽  
Migratory Cells

Cell surface heparan sulfate proteoglycans have been implicated as co-receptors facilitating cell adhesion and growth factor binding. Recent studies on the role of a family of transmembrane heparan sulfate proteoglycans, syndecans, in cell adhesion has identified one member, syndecan-4, to be present within focal contacts. The current study investigates the mechanisms regulating the association of syndecan-4 with focal contacts based upon its immunolocalization with vinculin in quiescent, serum-stimulated, and 12-0-tetradecanoylphorbol 13-acetate (TPA)-induced cultures. In quiescent cells, syndecan-4 did not localize to focal contacts. However, activation of protein kinase C by TPA or serum induces the active recruitment of syndecan-4 into focal contacts. This induction preferentially localizes syndecan-4 to focal contacts behind the leading lamella, the subnuclear region, and along the trailing edge of migratory cells. Focal contacts in either freshly adhered cells or in the leading lamellae of migrating cells did not stain for syndecan-4. In addition to the observed subcellular distribution and recruitment, syndecan-4 was observed to co-localize with endogenously synthesized fibronectin fibrils within focal contacts as well as with fibrils present in the matrix. These findings suggest that protein kinase C activation results in syndecan-4 recruitment to focal contacts and its association with sites of matrix deposition.

scholarly journals Requirement for diacylglycerol and protein kinase C in HeLa cell-substratum adhesion and their feedback amplification of arachidonic acid production for optimum cell spreading.

1993 ◽  
Vol 4 (3) ◽  
pp. 271-281 ◽  
Author(s):  
J S Chun ◽  
B S Jacobson
Keyword(s):  
Arachidonic Acid ◽  
Protein Kinase C ◽  
Protein Kinase ◽  
Hela Cell ◽  
Cell Attachment ◽  
Cell Spreading ◽  
Subsequent Formation ◽  
Sequence Of Events ◽  
Kinase C ◽  
Pkc Activation

Release of arachidonic acid (AA) and subsequent formation of a lipoxygenase (LOX) metabolite(s) is an obligatory signal to induce spreading of HeLa cells on a gelatin substratum (Chun and Jacobson, 1992). This study characterizes signaling pathways that follow the LOX metabolite(s) formation. Levels of diacylglycerol (DG) increase upon attachment and before cell spreading on a gelatin substratum. DG production and cell spreading are insignificant when phospholipase A2 (PLA2) or LOX is blocked. In contrast, when cells in suspension where PLA2 activity is not stimulated are treated with exogenous AA, DG production is turned on, and inhibition of LOX turns it off. This indicates that the formation of a LOX metabolite(s) from AA released during cell attachment induces the production of DG. Consistent with the DG production is the activation of protein kinase C (PKC) which, as with AA and DG, occurs upon attachment and before cell spreading. Inhibition of AA release and subsequent DG production blocks both PKC activation and cell spreading. Cell spreading is also blocked by the inhibition of PKC with calphostin C or sphingosine. The inhibition of cell spreading induced by blocking AA release is reversed by the direct activation of PKC with phorbol ester. However, the inhibition of cell spreading induced by PKC inhibition is not reversed by exogenously applied AA. In addition, inhibition of PKC does not block AA release and DG production. The data indicate that there is a sequence of events triggered by HeLa cell attachment to a gelatin substratum that leads to the initiation of cell spreading: AA release, a LOX metabolite(s) formation, DG production, and PKC activation. The data also provide evidence indicating that HeLa cell spreading is a cyclic feedback amplification process centered on the production of AA, which is the first messenger produced in the sequence of messengers initiating cell spreading. Both DG and PKC activity that are increased during HeLa cell attachment to a gelatin substratum appear to be involved. DG not only activates PKC, which is essential for cell spreading, but is also hydrolyzed to AA. PKC, which is initially activated as consequence of AA production, also increases more AA production by activating PLA2.

Protein Kinase C-Mediated Desmin Phosphorylation is Related to Myofibril Disarray in Cardiomyopathic Hamster Heart1

2002 ◽  
Vol 227 (11) ◽  
pp. 1039-1046 ◽  
Author(s):  
Xupei Huang ◽  
Jian Li ◽  
Dalton Foster ◽  
Sharon L. Lemanski ◽  
Dipak K. Dube ◽  
...  
Keyword(s):  
Protein Kinase C ◽  
Protein Kinase ◽  
Premature Death ◽  
Cardiac Cells ◽  
Immunogold Electron Microscopy ◽  
Intermediate Filament Protein ◽  
Kinase C ◽  
Hamster Strain ◽  
Significant Difference ◽  
Hereditary Cardiomyopathy

The cardiomyopathic (CM) Syrian golden hamster (strain UM-X7.1) exhibits a hereditary cardiomyopathy, which causes premature death resulting from congestive heart failure. The CM animals show extensive cardiac myofibril disarray and myocardial calcium overload. The present study has been undertaken to examine the role of desmin phosphorylation in myofibril disarray observed in CM hearts. The data from skinned myofibril protein phosphorylation assays have shown that desmin can be phosphorylated by protein kinase C (PKC). There is no significant difference in the content of desmin between CM and control hamster hearts. However, the desmin from CM hearts has a higher phosphorylation level than that of the normal hearts. Furthermore, we have examined the distribution of desmin and myofibril organization with immunofluorescent microscopy and immunogold electron microscopy in cultured cardiac myocytes after treatment with the PKC-activating phorbol ester, 12-O-tetradecanylphorbol-13-acetate (TPA). When the cultured normal hamster cardiac cells are treated with TPA, desmin filaments are disassembled and the myofibrils become disarrayed. The myofibril disarray closely mimics that observed in untreated CM cultures. These results suggest that disassembly of desmin filaments, which could be caused by PKC-mediated phosphorylation, may be a factor in myofibril disarray in cardiomyopathic cells and that the intermediate filament protein, desmin, plays an Important role in maintaining myofibril alignment in cardiac cells.

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