© 2000 American Heart Association, Inc.
Integrative Physiology |
Early Activation of the Multicomponent Signaling Complex Associated With Focal Adhesion Kinase Induced by Pressure Overload in the Rat Heart
From the Department of Internal Medicine, School of Medicine, State University of Campinas (UNICAMP), Campinas, São Paulo, Brazil.
Correspondence to Kleber G. Franchini, MD, Departamento de Clínica Médica, Faculdade de Ciências Médicas, Universidade Estadual de Campinas, Cidade Universitária "Zefferino Vaz," 13081-970 Campinas, São Paulo, Brasil. E-mail franchin{at}obelix.unicamp.br
 |
Abstract |
|---|
 Top Abstract IntroductionMaterials and MethodsResultsDiscussionReferences |
|---|
Abstract—Mechanical overload elicits functional and structural adaptive mechanisms in cardiac muscle. Signaling pathways linked to integrin/cytoskeleton complexes may have a function in mediation of the effects of mechanical stimulus in myocardial cells. We investigated the tyrosine phosphorylation and the assembly of the multicomponent signaling complex associated with focal adhesion kinase (Fak) and the actin cytoskeleton in the overloaded myocardium of rats. Pressure overload induced a 3-fold increase in Fak tyrosine phosphorylation within 3 minutes after a 60-mm Hg rise in aortic pressure. A pressure stimulus that lasted for 60 minutes was accompanied by a 5-fold increase in the amount of tyrosine-phosphorylated Fak, and a stimulus as low as 10 mm Hg doubled the amount of tyrosine-phosphorylated Fak in the myocardium within 10 minutes. Pressure overload also induced a time-dependent association of actin with Fak and an increase in the amount of Fak detected in the cytoskeletal fraction of the myocardium. These events were paralleled by c-Src activation and binding to Fak and by an association of Grb2 and p85 subunit of phosphatidylinositol 3-kinase with Fak. Erk1/2 and Akt, two possible downstream effectors of Fak via Grb2 and phosphatidylinositol 3-kinase, were also shown to be activated in parallel with Fak. These findings show that pressure overload induced a rapid activation of the Fak multiple signaling complex in the myocardium of rats, which suggests that this mechanism may have a role in mechanotransduction in the myocardium.
Key Words: myocardium • signaling • rats • kinase
|
Introduction |
|---|
|
TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
|---|
Mechanical stress is the major factor responsible for functional and structural adjustments of the myocardium in response to increased workload.1 Mechanosensitive ion channels and the release of autocrine and paracrine factors have been suggested to act as potential mechanisms that link mechanical stimuli to cellular responses in the myocardium.2 3 4
Mechanical stimuli may also trigger cellular signaling mechanisms
through the cytoskeleton via the elastic coupling to sites such as
plasma membrane, internal organelles, or nucleus.5 6 7 In
addition, the filamentous cytoskeletal network provides a scaffold
where signaling proteins can anchor and become involved in signal
transduction pathways.8 9 The activation of signaling
systems associated with cytoskeleton in tissues is fundamentally
dependent on the clustering of transmembrane integrins that act as
linkers between extracellular matrix proteins and the intracellular
cytoskeletal scaffold. Integrins connect to a meshwork of F-actin
through bridging proteins such as vinculin, talin, and
-actinin at specialized membrane-bound regions known as focal
adhesion complexes. These regions are rich in a variety of signaling
molecules, including focal adhesion kinase (Fak), c-Src family kinases,
guanine nucleotide exchange factors, Ras family proteins,
phosphatidylinositol 3 (PI3)-kinase, and mitogen-activated
protein kinases.8 9 After mechanical stimulation, integrin
clustering and engagement lead to a marked increase in tyrosine
phosphorylation and the recruitment of several cellular
proteins to the actin meshwork, in particular including
Fak.10 11 The precise mechanism that links integrin to Fak
activation is unknown, although it is clear that integrin clustering
mediates Fak autophosphorylation, predominantly at
Tyr397.12 After autophosphorylation,
additional tyrosine residues of Fak are phosphorylated
through the action of c-Src family kinases that bind to Fak at Tyr397
via their Src homology 2 (SH2) domains. This leads to the binding of
other SH2 domain proteins such as PI3 kinase13 and the
Grb2/Sos complex,14 which can then activate
signaling pathways that are involved in multiple cellular
processes.
Multiple integrins are expressed in the heart and may participate in biological processes such as intracellular pH regulation and hypertrophic growth.15 16 17 In addition, mechanical stimuli such as hypotonic cell swelling and pulsatile stretch have been shown to stimulate Fak tyrosine phosphorylation.18 19 In the myocardium of cats, a 4-hour period of pressure overload leads to tyrosine phosphorylation and activation of c-Src and its redistribution from cytosol to the cytoskeletal compartment.20 However, in this study, the authors were unable to show substantial levels of tyrosine phosphorylation of Fak.
Although this evidence suggests that the Fak signaling complex is activated by and contributes to the adaptive myocardial responses to mechanical stimuli, a clear demonstration of this relationship is still lacking. In the present study, we examined the phosphorylation and activation of the Fak signaling complex in the rat heart during acute pressure overload produced through controlled constriction of the transverse aorta. Moreover, possible downstream pathways mobilized by the Fak signaling complex were explored.
|
Materials and Methods |
|---|
|
TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
|---|
Antibodies and Chemicals
Polyclonal rabbit anti-Fak, Grb2, Erk1/2, and monoclonal mouse anti-phosphotyrosine, anti–phospho-Erk1/2 (Thr202/Tyr204), and anti–c-Src antibodies were obtained from Santa Cruz Biotechnology. Polyclonal rabbit anti–phospho-c-Src (Tyr416) was obtained from BioSource International, Inc. Polyclonal rabbit anti-p85 subunit of PI3 kinase was from Upstate Biotechnology. Polyclonal rabbit anti-Akt and anti–phospho-Akt (Ser473) were from New England Biolabs, Inc. Monoclonal mouse anti–nonsarcomeric
-actin was from Zymed.
Affinity-purified rabbit anti-mouse IgG was from DAKO.
[125I]Protein A and
[
-32P]ATP were from Amersham. Protein
A–Sepharose 6 MB was from Pharmacia. All other reagents were from
Sigma Chemical Co.
Animals
Adult male Wistar rats (n=60; weight 180 to 220 g) were
obtained from the animal facilities of the university. All procedures
and care of the rats were in accordance with institutional guidelines
for the use of animals in research.
Pressure overload was induced in pentobarbital-anesthetized
rats (50 mg/kg IP) through constriction of the transverse aorta with an
adjustable clamp. After anesthesia was induced, the animals
were maintained under controlled temperature and ventilation. The
aortic and vagus nerves were sectioned bilaterally to minimize the
influence of neural reflex on hemodynamics during
aortic constriction. The transverse aorta was accessed through the
second left intercostal space. A customized adjustable clamp was placed
around this vessel, after which the thoracic cavity was closed. After
stabilization (
20 minutes), pressure overload protocols were started
with adjustment of the aortic clamping while blood pressure signals
from above and below the constriction were monitored. The experimental
protocols included sustained (60 mm Hg; 3 to 60 minutes) and
stepwise (from 10 to 30 mm Hg; 10 minutes) increases in the
ascending aorta blood pressure. At the end of the blood
pressure–recording period, the ventricles were rapidly
removed, minced coarsely, and homogenized. Sham animals
were prepared in the same way except for the aortic constriction.
Protein Analysis With Immunoblotting
Aliquots of whole extracts or immunoprecipitated proteins that
contained equal amounts of total protein were treated with Laemmli’s
sample buffer and underwent SDS-PAGE. The nitrocellulose membranes with
transferred proteins were incubated with specific antibodies and
[125I]Protein A. Band intensities were
quantified through optical densitometry of the developed
autoradiographs.
Isolation of Cardiac Cytoskeleton
Cardiac cytoskeletal preparations were obtained from 100 mg
ventricular tissue as described previously.20
Samples of the cytoskeletal fraction and of the soluble fraction
obtained through ultracentrifugation underwent
SDS-PAGE, were transferred to nitrocellulose membranes, and were
blotted with anti-Fak antibody.
Immune Complex Tyrosine Kinase Reactions
Tyrosine kinase activity associated with the immune complex of
the anti-Fak antibody was assayed with denatured rabbit muscle enolase
used as substrate.21 After SDS-PAGE, the proteins were
transferred to nitrocellulose membranes, and the
32P-enolase band (46 kDa) was visualized and
quantified with autoradiography.
Statistical Analysis
The data are presented as mean±SEM. Differences between
the mean values of the densitometric readings were tested with 1-way
ANOVA for repeated measurements and Bonferroni’s multiple-range test.
A value of P<0.05 indicated statistical significance.
An expanded Materials and Methods section can be found in an online data supplement available at http://www.circresaha.org.
|
Results |
|---|
|
TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
|---|
Pressure Overload Induces Tyrosine Phosphorylation and Activation of p125Fak and p60Src
The tyrosine phosphorylation of Fak is closely related to its kinase activity.12 13 Figure 1
shows that acute pressure overload
increased the p125Fak phosphotyrosine content
mainly in the left ventricle, whereas a comparable amount of this
protein was found in both ventricles. Next, we studied
p125Fak tyrosine phosphorylation
induced with stepwise increases in aortic blood pressure (Figure 2). p125Fak
phosphotyrosine content increased to 180% in response to pressure
increases of 10 mm Hg. Additional increases were seen in hearts
when the constriction increased aortic pressure by 20 and 30
mm Hg (to 220% and 240%, respectively). The time course of
p125Fak tyrosine phosphorylation
was examined in hearts subjected to sustained increases in aortic
pressure of 60 mm Hg (Figure 3).
p125Fak phosphotyrosine content increased 3-fold
within 3 minutes and 5-fold within 60 minutes of continuous and stable
pressure increases. Parallel immunoblots with anti-Fak
antibody revealed that the amount of p125Fak in
the myocardium remained unaltered during this period.
|
|
|
After activation, Fak autophosphorylates Tyr397, creating a
binding site for SH2 domain of c-Src. c-Src kinase is then
activated with phosphorylation on
Tyr416.12 The activation of c-Src also depends on the
dephosphorylation of constitutively
phosphorylated Tyr527.12
Coimmunoprecipitation assays with anti-Fak and anti–c-Src antibodies
showed only a weak binding of p60c-Src to Fak in
the myocardium of control rats (Figure 4A). Pressure overload increased the
p60c-Src binding to Fak, in parallel with its
tyrosine phosphorylation. We next examined whether
pressure overload also induces tyrosine phosphorylation
and activation of p60c-Src. Immunoprecipitation
experiments with anti–c-Src antibodies showed that the tyrosine
phosphorylation of p60c-Src
increased in overloaded compared with unloaded hearts (Figure 4B). This finding could imply either activation or inactivation,
depending on whether the phosphorylation was at Tyr416
or Tyr527.12 We then examined the c-Src tyrosine
phosphorylation with a phosphospecific antibody against
c-Src (Tyr416). As demonstrated in Figure 4C, pressure overload
induced a rapid and sustained increase in phospho-c-Src (Tyr416),
indicating that p60c-Src is activated in
parallel with Fak. This occurred while the amount of
p60c-Src remained constant (Figure 4D).
|
Further indication of load-induced Fak/c-Src association and c-Src
activation was provided by experiments in which the kinase activity of
the immune complex of anti-Fak antibody was tested against rabbit
denatured muscle enolase, a substrate for c-Src but not for
Fak.21 Figure 5 shows that
there was negligible tyrosine kinase activity in the immune complex of
anti-Fak antibody in unloaded hearts. However, the enolase
phosphorylation increased greatly (to 150% after 3
minutes and to 350% after 60 minutes of pressure overload) when
incubated with immunoprecipitates of overloaded hearts.
|
Redistribution of p125Fak to the Cytoskeletal
Compartment Induced by Pressure Overload
Because the migration of Fak to the actin meshwork is a crucial
event for Fak activation,8 9 we performed experiments to
detect the association of actin with p125Fak. The
membranes used to detect p125Fak were stripped
and blotted with antibody against nonsarcomeric -actin. As shown in
Figure 6A, overloaded hearts showed a
time-dependent coimmunoprecipitation of actin with
p125Fak. The pressure-induced migration of
p125Fak to the cytoskeletal compartment was
further suggested by experiments with cytoskeletal fraction
preparations obtained with differential centrifugation.
A substantial increase in the amount of p125Fak
was detected in the cytoskeletal fraction of homogenates
from overloaded myocardial tissue (Figure 6B).
|
Pressure Overload Induces the Association of Fak With Grb2
and Activation of ERK1/2
Phosphorylation of Fak Tyr925 by c-Src confers a
site for Grb2 binding, which potentially links integrin/Fak signaling
to the Ras/mitogen-activated protein kinase
pathway.14 As shown in Figure 7A, acute pressure overload increased the
amount of Grb2 binding to Fak in parallel with activation of the
Fak/c-Src complex.
|
We next investigated whether activation of the Fak/c-Src complex was
paralleled by the activation of Erk1/2, a possible downstream
effector of Fak via Grb2 binding.21 Western blotting with
anti–phospho-Erk1/2 (Thr202/Tyr204)–specific antibody revealed an
increase and presumably an activation of this enzyme within 5 minutes
(to 180%), with a maximal activation being achieved by 10 minutes
(to 240%) of pressure overload. The values of phospho-Erk1/2
returned toward the baseline values after 60 minutes of pressure
overload (Figure 7B
). Western blotting with an antibody for
Erk1/2 that detects both the phosphorylated and
unphosphorylated forms of the kinases showed that
myocardial Erk1/2 levels were similar in the heart studied at various
periods after aortic constriction (Figure 7C).
Pressure Overload Induces the Association of Fak With p85
Subunit of PI3 Kinase and Activation of Akt
In addition of being a site for c-Src binding, Tyr397 of Fak
has been identified as the major site for binding of PI3 kinase, whose
inositol lipid products are key mediators of multiple intracellular
signaling.13 22 As shown in Figure 8A, pressure overload induced a rapid
increase in the association of Fak with p85 subunit of PI3 kinase (to
180% within 3 minutes), increasing to 360% within 1 hour of
aortic constriction. These results indicated that pressure overload
induced a recruitment and a possible activation of this enzyme.
|
One of the multiple downstream signaling molecules regulated by
3'-phosphorylated phosphatidylinositides is the
serine/threonine protein kinase Akt.22 In many cases, the
activation of Akt is initiated by the binding of 3
phosphoinositides to its pleckstrin homology domain,
translocation from the cytoplasm to the plasma membrane, and subsequent
phosphorylation by upstream kinases, including PDK1. In
the present study, we also examined whether pressure overload
activates Akt. Western blotting with anti-Akt
(Ser473)-phosphospecific antibody revealed an increase and presumably
an activation of this enzyme within 3 minutes (to 200%) after the
beginning of pressure overload stimulus. The amount of phospho-Akt
(Ser473) remained increased to 300% up to 1 hour of pressure
overload (Figure 8B), whereas the amount of Akt remained
unchanged during this period (Figure 8C).
|
Discussion |
|---|
|
TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
|---|
The data presented here support the conclusion that load induces a rapid assembly and activation of the multicomponent signaling complex associated with Fak in the rat heart. The close relationship between increased load and Fak/c-Src activation and the concurrent activation of Erk1/2 and Akt, two potential downstream effectors of the Fak multicomponent signaling complex, indicated that it may play a role in the earlier myocardial responses to increased workload.
Load-Induced Activation and Assembly of the Fak Multicomponent
Signaling Complex
Pressure overload induced a rapid increase (3 minutes) in the
myocardial p125Fak phosphotyrosine content,
preferentially in the left ventricle. Further increases were observed
after 1 hour of augmented workload. This was accompanied by a
load-induced association of c-Src with Fak, as indicated by the
increase in the amount of p60c-Src detected in
blots of immunoprecipitated Fak, and c-Src activation, as indicated by
the increase in the amount of c-Src detected with phosphospecific
antibody against c-Src (Tyr416) in overloaded compared with unloaded
hearts. The idea that load induces a rapid activation of c-Src and its
association with Fak was strengthened by the increased kinase activity
detected in the immune complex of anti-Fak antibody toward the c-Src
substrate enolase. Our study also provides evidence that load induces
the association of the signaling molecules Grb2 and PI3 kinase to Fak
and the migration of p125Fak to the cytoskeletal
compartment. Presumably, this p125Fak migration
was directed to the actin meshwork, as suggested by the time-dependent
association of actin with Fak in overloaded hearts.
These results are in general consistent with data from noncardiac cells that demonstrate that mechanical stimulus activated the Fak/c-Src complex and its recruitment to the actin cytoskeletal meshwork, presumably at sites related to focal adhesion.8 9 Cardiac myocytes contain structures known as costameres, which resemble focal adhesion complexes and have been suggested to be involved in the transduction of mechanical forces from cardiac myocyte surface.23 24 Costameres are also rich in vinculin, talin, integrins, and a meshwork of actin that occur in register with Z lines.23 24 However, a clear demonstration that Fak is localized or migrates to costameres on mechanical stimuli must be confirmed.
The clear relationship between the assembly and activation of the Fak multicomponent signaling complex with the load stimuli suggests that this mechanism may play a central role in the mechanotransduction during increased load in the myocardium. In this context, studies in cultured rat cardiomyocytes have shown that pulsatile stretch can activate Fak within 5 minutes after the beginning of the stretch.19 As has been shown in cultured cells,10 it is possible that this rapid activation of the Fak/c-Src signaling complex is mediated via integrin, but Fak tyrosine phosphorylation and activation may also be elicited through a variety of nonintegrin cell surface receptors, including growth factor tyrosine kinases and G protein–coupled receptors.9 However, Fak activation mediated by soluble factors has been shown to be short in duration, which favors the hypothesis that the activation of Fak seen in the present study is predominantly mediated via integrins.8 9
In contrast with the present results, previous studies20 have failed to detect substantial phosphorylation of Fak in pressure-overloaded myocardium. One possible explanation for this discrepancy may be related to the fact that in previous studies, the tyrosine phosphorylation of Fak was examined after 4 hours of pressure overload, by which time the tyrosine phosphorylation of Fak may have vanished because of the actions of tyrosine phosphatases. However, this explanation is not consistent with data that show a persistent activation of nonreceptor tyrosine kinases for longer periods in overloaded myocardium.20 Experiments with controlled aortic constriction for longer periods could clarify the difference between the results of the present study and those of previous studies. However, the maintenance of a stable and reliable preparation with aortic constriction for periods of >1 hour is difficult in anesthetized rats.
Load-Induced Activation of ERK1/2 and Akt
The activation of Fak has been shown to influence a variety of
cellular functions, including the control of cell shape, growth, and
survival.8 9 In looking at possible downstream events
regulated by the Fak signaling complex, we also showed that Erk1/2 and
Akt were activated in parallel with activation of the Fak
signaling complex. The rapid association of Fak with Grb2 and PI3
kinase, two intermediate steps between Fak and Erk1/2 and Akt
activation, strengthened the idea that Fak/c-Src activation may be the
upstream event of such a rapid activation of these kinases, as has been
suggested previously.21 25
Erk1/2 regulates an extensive range of cellular processes, including
gene transcription, cytoskeletal organization, metabolic
homeostasis, cell growth, and survival.26 The activation
of Erk1/2 is likely to be an important pathway to the adaptation of
myocardial cells to mechanical stimuli. To date, mechanical stretch of
neonatal cardiac myocytes has been shown to rapidly (within 5 minutes)
activate the Ras/Erk1/2 pathway.27 28 Although the
functional roles of these Erks in stretch-induced cardiac
hypertrophy are unclear at the present, they may be
important in the regulation of the expression of genes such as early
responsive genes (eg, c-fos, c-jun,
egr-1), the expression of which is rapidly and transiently
upregulated in the myocardium and isolated
cardiomyocytes subjected to mechanical stress. Moreover,
the early activation of Erk1/2 has been suggested to contribute to the
reexpression of fetal ventricle genes (eg, atrial
natriuretic factor, ß-myosin heavy chain, and skeletal
muscle -actin).29
Akt activation transduces signals that regulate multiple biological processes, including glucose metabolism, apoptosis, gene expression, and cell proliferation.22 Studies have shown that Akt mediates the antiapoptosis effects of Fak.25 In cardiac myocytes, it has been shown that Akt mediates ß-adrenergic receptor–stimulated atrial natriuretic factor transcription.30 To our knowledge, the present study is the first to show that Akt may be activated during the early response to pressure overload in the myocardium. The importance of the early activation of Akt during pressure overload can only be speculated. Fak-mediated activation of Akt is likely to be important for the overall response to increased load. Akt phosphorylates various intracellular substrates, thereby affecting metabolism,31 protein synthesis,32 cell survival/apoptosis,33 and gene expression through the regulation of transcription factors,34 which could mediate the cellular responses to increased load in the myocardium.
The present results do not exclude the possibility that mechanisms other than Fak/Src activation could be responsible for the activation of Erk1/2 and Akt during the early phase of cardiac response to pressure overload. To date, Erks and Akt have been shown to be activated through protein–tyrosine kinase and G protein–coupled receptors.22 26 Therefore, further studies are necessary to confirm the importance of the Fak signaling complex in the activation of Erk and Akt.
In conclusion, pressure overload induced a rapid assembly of the Fak/Src signaling complex in the myocardium. This activation was shown to be roughly parallel to stimulus intensity and duration and to the activation of possible downstream pathways such as Erk1/2 and Akt activation. In consideration of the potential effects of this signaling system on multiple cellular functions such as ion transport, force transmission, metabolic pathways, intracellular molecular transport, survival, and gene expression, this complex may occupy a central position in the adaptive changes induced by increased load in the myocardium.
|
Acknowledgments |
|---|
This study was sponsored by grants from Fundação de Auxílio à Pesquisa do Estado de São Paulo (FAPESP; Proc. 98/11403-7) and Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq; Proc. 521098/97-1).
Received March 10, 2000; revision received July 10, 2000; accepted July 28, 2000.
|
References |
|---|
|
TopAbstractIntroductionMaterials and MethodsResultsDiscussionReferences |
|---|
1. Cooper G. Basic determinants of myocardial hypertrophy: a review of molecular mechanisms. Annu Rev Med. 1997;48:13–23.[Medline] [Order article via Infotrieve]
2.
Kent RL, Hoober JK, Cooper G. Load responsiveness of
protein synthesis in adult mammalian myocardium: role of
cardiac deformation linked to sodium influx. Circ Res. 1989;64:74–85.
3. Kaye D, Pimental D, Prasad S, Maki T, Berger HJ, McNeil PL, Smith TW, Kelly RA. Role of transient sarcolemmal membrane permeability and basic fibroblast growth factor release in the hypertrophic response of adult rat ventricular myocytes to increased mechanical activity in vitro. J Clin Invest. 1996;97:281–291.[Medline] [Order article via Infotrieve]
4. Sadoshima J-I, Xu Y, Slayter HS, Izumo S. Autocrine release of angiotensin II mediates stretch-induced hypertrophy of cardiac myocytes in vitro. Cell. 1993;75:977–984.[Medline] [Order article via Infotrieve]
5. Ingber D. Tensegrity: the architectural basis of cellular mechano-transduction. Annu Rev Physiol. 1997;59:575–599.[Medline] [Order article via Infotrieve]
6.
Janmey PA. The cytoskeleton and cell signaling:
component localization and mechanical coupling. Physiol Rev. 1998;78:763–781.
7.
Maniotis AJ, Chen CS, Ingber DE. Demonstration of
mechanical connections between integrins, cytoskeletal filaments, and
nucleoplasm that stabilize nuclear structure. Proc Natl Acad
Sci. 1997;94:849–854.
8. Burridge K, Chrzanowska-Wodnicka M. Focal adhesions, contractility, and signaling. Annu Rev Cell Dev Biol. 1996;12:463–519.[Medline] [Order article via Infotrieve]
9.
Giancotti FG, Ruoslahti E. Integrin signaling.
Science. 1999;285:1028–1032.
10.
Burridge K, Turner CE, Romer LH. Tyrosine
phosphorylation of paxillin and pp125 FAK accompanies
cell adhesion to extracellular matrix: a role in cytoskeletal assembly.
J Cell Biol. 1992;119:893–903.
11.
Kornberg L, Earp HS, Parsons JT, Schaller M, Juliano
RL. Cell adhesion or integrin clustering increases
phosphorylation of a focal adhesion-associated tyrosine
kinase. J Biol Chem. 1992;267:23439–23442.
12.
Schaller MD, Hildebrand JD, Shannon JD, Fox JW, Vines
RR, Parsons JT. Autophosphorylation of the focal
adhesion kinase, pp125FAK, directs SH2-dependent binding of pp60src.
Mol Cell Biol. 1994;14:1680–1688.
13. Bachelot C, Rameh L, Parsons T, Cantley LC. Association of phosphatidylinositol 3-kinase, via SH2 domains of p85, with focal adhesion kinase in polyoma middle t-transformed fibroblasts. Biochim Biophys Acta. 1996;1311:45–52.[Medline] [Order article via Infotrieve]
14. Schlaepfer DD, Hanks S, Hunter T, van der Geer P. Integrin-mediated signal transduction linked to Ras pathway by GRB2 binding to focal adhesion kinase. Nature. 1994;372:786–791.[Medline] [Order article via Infotrieve]
15.
Terracio L, Rubin K, Gullberg D, Balog E, Carver W,
Jyring R, et al. Expression of collagen-binding integrins during
cardiac development and hypertrophy. Circ Res. 1991;68:734–744.
16.
Pucéat M, Roche S, Vassort G. Src family tyrosine
kinase regulates intracellular pH in cardiomyocytes.
J Cell Biol. 1998;141:1637–1646.
17.
Ross RS, Pham C, Shai SY, Goldhaber JI, Fenczik C,
Glembotski CC, Ginsberg MH, Loftus JC. ß1
Integrin participates in the hypertrophic response of rat
ventricular myocytes. Circ Res. 1998;82:1160–1172.
18. Sadoshima J, Izumo S. Roles of integrins in cell swelling-induced tyrosine phosphorylation in cardiac myocytes. Circulation. 1994;90(suppl I):I-305. Abstract.
19. Seko Y, Takahashi N, Tobe K, Kadowaki T, Yazaki Y. Pulsatile stretch activates mitogen-activated protein kinase (MAPK) family members and focal adhesion kinase (p125(FAK)) in cultured rat cardiac myocytes. Biochem Biophys Res Commun. 1999;259:8–14.[Medline] [Order article via Infotrieve]
20.
Kuppuswamy D, Kerr C, Narishige T, Kasi VS, Menick DR,
Cooper G. Association of tyrosine-phosphorylated c-Src
with the cytoskeleton of hypertrophying
myocardium. J Biol Chem. 1997;272:4500–4508.
21.
Schlaepfer DD, Hunter T. Focal adhesion kinase
overexpression enhances Ras-dependent integrin signaling to
Erk2/mitogen-activated protein kinase through interactions with
and activation of c-Src. J Biol Chem. 1997;272:13189–13195.
22. Chan TO, Rittenhouse SE, Tsichlis PN. Akt/PKB and other D3 phosphoinositides-regulated kinases: kinase activation by phosphoinositide-dependent phosphorylation. Annu Rev Biochem. 1999;68:965–1014.[Medline] [Order article via Infotrieve]
23.
Pardo JV, Siciliano JD, Craig SW. Vinculin is a
component of an extensive network of myofibril-sarcolemma attachment
regions in cardiac muscle fibers. J Cell Biol. 1983;97:1081–1088.
24.
Danowski BA, Imanaka-Yoshida K, Sanger JM, Sanger JW.
Costameres are sites of force transmission to the substratum in adult
rat cardiomyocytes. J Cell Biol. 1992;118:1411–1420.
25.
Sonoda Y, Watanabe S, Matsumoto Y, Aizu-Yokota E,
Kasahara T. FAK is the upstream signal protein of the
phosphatidylinositol 3-kinase-Akt survival pathway in hydrogen
peroxide-induced apoptosis of a human glioblastoma cell line.
J Biol Chem. 1999;274:36544–36549.
26. Seger R, Krebs EG. The MAPK signaling cascade. FASEB J. 1995;9:726–735.[Abstract]
27. Sadoshima J, Izumo S. Mechanical stretch rapidly activates multiple signal transduction pathways in cardiac myocytes: potential involvement of an autocrine/paracrine mechanism. EMBO J. 1993;12:1681–1692.
28.
Yamazaki T, Tobe K, Hoh E, Maemura K, Kaida T, Komuro
I, Tamemoto H, Kadowaki T, Nagai R, Yazaki Y. Mechanical loading
activates mitogen-activated protein kinase and S6
peptide kinase in cultured rat cardiac myocytes. J Biol
Chem. 1993;268:12069–12076.
29.
Thorburn J, Frost JA, Thorburn A.
Mitogen-activated protein kinases mediate changes in gene
expression, but not cytoskeletal organization associated with cardiac
muscle cell hypertrophy. J Cell Biol. 1994;126:1565–1572.
30.
Morisco C, Zebrowski D, Condorelli G, Tsichlis P,
Vatner SF, Sadoshima J. The Akt-glycogen synthase kinase 3ß
pathway regulates transcription of atrial natriuretic
factor induced by ß-adrenergic receptor stimulation in cardiac
myocytes. J Biol Chem. 2000;275:14466–14475.
31. Cross DA, Alessi DR, Cohen P, Andjelkovich M, Hemmings BA. Inhibition of glycogen synthase kinase-3 by insulin mediated by protein kinase B. Nature. 1995;378:785–789.[Medline] [Order article via Infotrieve]
32. Burgering BM, Coffer PJ. Protein kinase B (c-Akt) in phosphatidylinositol-3-OH kinase signal transduction. Nature. 1995;376:599–602.[Medline] [Order article via Infotrieve]
33. Brennan P, Babbage JW, Burgering BM, Groner B, Reif K, Cantrell DA. Phosphatidylinositol 3-kinase couples the interleukin-2 receptor to the cell cycle regulator E2F. Immunity. 1997;7:679–689.[Medline] [Order article via Infotrieve]
34. Downward J. Mechanisms and consequences of activation of protein kinase B/Akt. Curr Opin Cell Biol. 1998;10:262–267.[Medline] [Order article via Infotrieve]
This article has been cited by other articles:
 |
|
 |
|
  D. P. Del Re, S. Miyamoto, and J. H. Brown Focal Adhesion Kinase as a RhoA-activable Signaling Scaffold Mediating Akt Activation and Cardiomyocyte Protection J. Biol. Chem., December 19, 2008; 283(51): 35622 - 35629. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 T. M. Marin, C. F.M.Z. Clemente, A. M. Santos, P. K. Picardi, V. D.B. Pascoal, I. Lopes-Cendes, M. J.A. Saad, and K. G. Franchini Shp2 Negatively Regulates Growth in Cardiomyocytes by Controlling Focal Adhesion Kinase/Src and mTOR Pathways Circ. Res., October 10, 2008; 103(8): 813 - 824. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
A. Sabri, K. Rafiq, R. Seqqat, M. A. Kolpakov, R. Dillon, and L. J. Dell'italia Sympathetic Activation Causes Focal Adhesion Signaling Alteration in Early Compensated Volume Overload Attributable to Isolated Mitral Regurgitation in the Dog Circ. Res., May 9, 2008; 102(9): 1127 - 1136. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 V. Thamilselvan, D. H. Craig, and M. D. Basson FAK association with multiple signal proteins mediates pressure-induced colon cancer cell adhesion via a Src-dependent PI3K/Akt pathway FASEB J, June 1, 2007; 21(8): 1730 - 1741. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
L. A. DiMichele, J. T. Doherty, M. Rojas, H. E. Beggs, L. F. Reichardt, C. P. Mack, and J. M. Taylor Myocyte-Restricted Focal Adhesion Kinase Deletion Attenuates Pressure Overload-Induced Hypertrophy Circ. Res., September 15, 2006; 99(6): 636 - 645. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 D. Srivastava and S. Yu Stretching to meet needs: integrin-linked kinase and the cardiac pump Genes & Dev., September 1, 2006; 20(17): 2327 - 2331. [Full Text] [PDF]
|
|
|
|
 |
|
 T. A. Baudino, W. Carver, W. Giles, and T. K. Borg Cardiac fibroblasts: friend or foe? Am J Physiol Heart Circ Physiol, September 1, 2006; 291(3): H1015 - H1026. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
X. P. Yi, J. Zhou, L. Huber, J. Qu, X. Wang, A. M. Gerdes, and F. Li Nuclear compartmentalization of FAK and FRNK in cardiac myocytes Am J Physiol Heart Circ Physiol, June 1, 2006; 290(6): H2509 - H2515. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 M. Brancaccio, E. Hirsch, A. Notte, G. Selvetella, G. Lembo, and G. Tarone Integrin signalling: The tug-of-war in heart hypertrophy Cardiovasc Res, June 1, 2006; 70(3): 422 - 433. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. Hoshijima Mechanical stress-strain sensors embedded in cardiac cytoskeleton: Z disk, titin, and associated structures Am J Physiol Heart Circ Physiol, April 1, 2006; 290(4): H1313 - H1325. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 D. M. Browe and C. M. Baumgarten EGFR Kinase Regulates Volume-sensitive Chloride Current Elicited by Integrin Stretch via PI-3K and NADPH Oxidase in Ventricular Myocytes J. Gen. Physiol., February 27, 2006; 127(3): 237 - 251. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 B. Han, M. Lodyga, and M. Liu Ventilator-induced Lung Injury: Role of Protein-Protein Interaction in Mechanosensation Proceedings of the ATS, October 1, 2005; 2(3): 181 - 187. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
W. Nadruz Jr., M. A.F. Corat, T. M. Marin, G. A. Guimaraes Pereira, and K. G. Franchini Focal adhesion kinase mediates MEF2 and c-Jun activation by stretch: Role in the activation of the cardiac hypertrophic genetic program Cardiovasc Res, October 1, 2005; 68(1): 87 - 97. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
A. S. Torsoni, T. M. Marin, L. A. Velloso, and K. G. Franchini RhoA/ROCK signaling is critical to FAK activation by cyclic stretch in cardiac myocytes Am J Physiol Heart Circ Physiol, October 1, 2005; 289(4): H1488 - H1496. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
P. M. Fonseca, R. Y. Inoue, C. B. Kobarg, D. P. Crosara-Alberto, J. Kobarg, and K. G. Franchini Targeting to C-Terminal Myosin Heavy Chain May Explain Mechanotransduction Involving Focal Adhesion Kinase in Cardiac Myocytes Circ. Res., January 7, 2005; 96(1): 73 - 81. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
P. H. Sugden Ras, Akt, and Mechanotransduction in the Cardiac Myocyte Circ. Res., December 12, 2003; 93(12): 1179 - 1192. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
D. M. Browe and C. M. Baumgarten Stretch of {beta}1 Integrin Activates an Outwardly Rectifying Chloride Current via FAK and Src in Rabbit Ventricular Myocytes J. Gen. Physiol., November 24, 2003; 122(6): 689 - 702. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. C. Heidkamp, A. L. Bayer, B. T. Scully, D. M. Eble, and A. M. Samarel Activation of focal adhesion kinase by protein kinase C{epsilon} in neonatal rat ventricular myocytes Am J Physiol Heart Circ Physiol, October 1, 2003; 285(4): H1684 - H1696. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
A. S. Torsoni, S. S. Constancio, W. Nadruz Jr, S. K. Hanks, and K. G. Franchini Focal Adhesion Kinase Is Activated and Mediates the Early Hypertrophic Response to Stretch in Cardiac Myocytes Circ. Res., July 25, 2003; 93(2): 140 - 147. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 A. S. Torsoni, P. M. Fonseca, D. P Crosara-Alberto, and K. G. Franchini Early activation of p160ROCK by pressure overload in rat heart Am J Physiol Cell Physiol, June 1, 2003; 284(6): C1411 - C1419. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 X. P. Yi, X. Wang, A. M. Gerdes, and F. Li Subcellular Redistribution of Focal Adhesion Kinase and Its Related Nonkinase in Hypertrophic Myocardium Hypertension, June 1, 2003; 41(6): 1317 - 1323. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J. Melendez, S. Welch, E. Schaefer, C. S. Moravec, S. Avraham, H. Avraham, and M. A. Sussman Activation of pyk2/Related Focal Adhesion Tyrosine Kinase and Focal Adhesion Kinase in Cardiac Remodeling J. Biol. Chem., November 15, 2002; 277(47): 45203 - 45210. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
D. E. Ingber Mechanical Signaling and the Cellular Response to Extracellular Matrix in Angiogenesis and Cardiovascular Physiology Circ. Res., November 15, 2002; 91(10): 877 - 887. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. A. Sussman, A. McCulloch, and T. K. Borg Dance Band on the Titanic: Biomechanical Signaling in Cardiac Hypertrophy Circ. Res., November 15, 2002; 91(10): 888 - 898. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 M. Lodyga, X.-H. Bai, E. Mourgeon, B. Han, S. Keshavjee, and M. Liu Molecular cloning of actin filament-associated protein: a putative adaptor in stretch-induced Src activation Am J Physiol Lung Cell Mol Physiol, August 1, 2002; 283(2): L265 - L274. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
A. L. Bayer, M. C. Heidkamp, N. Patel, M. J. Porter, S. J. Engman, and A. M. Samarel PYK2 expression and phosphorylation increases in pressure overload-induced left ventricular hypertrophy Am J Physiol Heart Circ Physiol, August 1, 2002; 283(2): H695 - H706. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. C. Heidkamp, A. L. Bayer, J. A. Kalina, D. M. Eble, and A. M. Samarel GFP-FRNK Disrupts Focal Adhesions and Induces Anoikis in Neonatal Rat Ventricular Myocytes Circ. Res., June 28, 2002; 90(12): 1282 - 1289. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
L. C. Hool Can Integrins Integrate Vascular Myogenic Responses? Circ. Res., March 8, 2002; 90(4): 371 - 373. [Full Text] [PDF]
|
|
|
|
|
|
P. P. Domingos, P. M. Fonseca, W. Nadruz Jr., and K. G. Franchini Load-induced focal adhesion kinase activation in the myocardium: role of stretch and contractile activity Am J Physiol Heart Circ Physiol, February 1, 2002; 282(2): H556 - H564. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
R. S. Ross and T. K. Borg Integrins and the Myocardium Circ. Res., June 8, 2001; 88(11): 1112 - 1119. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. J. Davis, X. Wu, T. R. Nurkiewicz, J. Kawasaki, G. E. Davis, M. A. Hill, and G. A. Meininger Integrins and mechanotransduction of the vascular myogenic response Am J Physiol Heart Circ Physiol, April 1, 2001; 280(4): H1427 - H1433. [Abstract] [Full Text] [PDF]
|
|
| ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||