© 2000 American Heart Association, Inc.
Cellular Biology |
The 
Subtype of Protein Kinase C Is Required for Cardiomyocyte Connexin-43 Phosphorylation
From the Institute of Cardiovascular Sciences (B.W.D., E.K.), University of Manitoba, St. Boniface General Hospital Research Centre, Winnipeg, Manitoba, Canada, and University of Louisville and Jewish Hospital Heart and Lung Institute (P.P.), Louisville, Ky.
Correspondence to E. Kardami, Institute of Cardiovascular Sciences, University of Manitoba, St Boniface General Hospital Research Centre, 351 Taché Ave, Winnipeg, MB, Canada, R2H 2A6. E-mail ekardami{at}sbrc.umanitoba.ca
Abstract—Gap junctions (GJs),
composed of connexins, are intercellular channels ensuring electric and
metabolic coupling between cardiomyocytes. We
have shown previously that an endogenous
mitogenic and cardioprotective protein, fibroblast growth
factor-2 (FGF-2), decreases cardiomyocyte GJ permeability
by stimulating phosphorylation of connexin-43 (Cx43).
Identifying the kinase(s) phosphorylating cardiac Cx43 may thus provide
a way of modulating cardiac intercellular communication. Because FGF-2
activates receptors linked to protein kinase C (PKC) and
mitogen-activated protein kinase, we first investigated
participation of these enzymatic systems in Cx43
phosphorylation. The inhibitor PD98059
blocked activation of mitogen-activated protein kinase, but it
did not prevent the FGF-2 effects on GJs. In contrast, the PKC
inhibitor chelerythrine blocked the effects of FGF-2 on
Cx43 phosphorylation and permeability. Because the
-isoform of PKC localizes to plasma membrane sites, we examined
whether it is directly involved in the FGF-2–induced Cx43
phosphorylation. In nonstimulated myocytes, PKC
displayed a discontinuous pattern of localization at intercellular
contact sites and partial colocalization with Cx43. Treatment with
FGF-2 or phorbol 12-myristate 13-acetate induced a more
continuous pattern of PKC distribution, whereas the anti-Cx43
staining appeared to overlap extensively with that of PKC. In
immunoprecipitation experiments using specific anti-Cx43 antibodies,
PKC but not PKC
coprecipitated with Cx43. FGF-2 increased levels
of coprecipitated PKC, suggesting increased association between
PKC and Cx43 on stimulation. Transient gene transfer and
overexpression of cDNAs coding for truncated or mutated
dominant-negative forms of PKC decreased cardiomyocyte
Cx43 phosphorylation significantly. We conclude that
PKC mediates the FGF-2–induced effects on cardiac GJs and that PKC
likely interacts with and phosphorylates cardiac Cx43 at
sites of intercellular contact.
Key Words: cardiomyocyte gap junction • protein kinase C • fibroblast growth factor-2 • phosphorylation
This article has been cited by other articles:
 |
|
 |
|
  S. F. Steinberg Structural Basis of Protein Kinase C Isoform Function Physiol Rev, October 1, 2008; 88(4): 1341 - 1378. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 A. Salameh, S. Krautblatter, S. Baessler, S. Karl, D. R. Gomez, S. Dhein, and D. Pfeiffer Signal Transduction and Transcriptional Control of Cardiac Connexin43 Up-Regulation after {alpha}1-Adrenoceptor Stimulation J. Pharmacol. Exp. Ther., July 1, 2008; 326(1): 315 - 322. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 S. Y. Lu, D. P. Sontag, K. A. Detillieux, and P. A. Cattini FGF-16 is released from neonatal cardiac myocytes and alters growth-related signaling: a possible role in postnatal development Am J Physiol Cell Physiol, May 1, 2008; 294(5): C1242 - C1249. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 M.-A. Meilleur, C. D. Akpovi, R.-M. Pelletier, and M. L. Vitale Tumor Necrosis Factor-{alpha}-Induced Anterior Pituitary Folliculostellate TtT/GF Cell Uncoupling Is Mediated by Connexin 43 Dephosphorylation Endocrinology, December 1, 2007; 148(12): 5913 - 5924. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 T. Miura, T. Yano, K. Naitoh, M. Nishihara, T. Miki, M. Tanno, and K. Shimamoto {delta}-Opioid receptor activation before ischemia reduces gap junction permeability in ischemic myocardium by PKC-{varepsilon}-mediated phosphorylation of connexin 43 Am J Physiol Heart Circ Physiol, September 1, 2007; 293(3): H1425 - H1431. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
V. Drobic, R. H. Cunnington, K. M. Bedosky, J. E. Raizman, V. V. Elimban, S. G. Rattan, and I. M. C. Dixon Differential and combined effects of cardiotrophin-1 and TGF-beta1 on cardiac myofibroblast proliferation and contraction Am J Physiol Heart Circ Physiol, August 1, 2007; 293(2): H1053 - H1064. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S. L. House, S. J. Melhorn, G. Newman, T. Doetschman, and J. E. J. Schultz The protein kinase C pathway mediates cardioprotection induced by cardiac-specific overexpression of fibroblast growth factor-2 Am J Physiol Heart Circ Physiol, July 1, 2007; 293(1): H354 - H365. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
K. Boengler, I. Konietzka, A. Buechert, Y. Heinen, D. Garcia-Dorado, G. Heusch, and R. Schulz Loss of ischemic preconditioning's cardioprotection in aged mouse hearts is associated with reduced gap junctional and mitochondrial levels of connexin 43 Am J Physiol Heart Circ Physiol, April 1, 2007; 292(4): H1764 - H1769. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 K. Naitoh, Y. Ichikawa, T. Miura, Y. Nakamura, T. Miki, Y. Ikeda, H. Kobayashi, M. Nishihara, K. Ohori, and K. Shimamoto MitoKATP channel activation suppresses gap junction permeability in the ischemic myocardium by an ERK-dependent mechanism Cardiovasc Res, May 1, 2006; 70(2): 374 - 383. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 S. Matsushita, H. Kurihara, M. Watanabe, T. Okada, T. Sakai, and A. Amano Alterations of Phosphorylation State of Connexin 43 during Hypoxia and Reoxygenation Are Associated with Cardiac Function J. Histochem. Cytochem., March 1, 2006; 54(3): 343 - 353. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 D. Chung and R. L. Caruso 2,2'-Dichlorobiphenyl Decreases Amplitude and Synchronization of Uterine Contractions Through MAPK1-Mediated Phosphorylation of GJA1 (Connexin43) and Inhibition of Myometrial Gap Junctions Biol Reprod, November 1, 2005; 73(5): 974 - 982. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S. Dhein New, emerging roles for cardiac connexins. Mitochondrial Cx43 raises new questions Cardiovasc Res, August 1, 2005; 67(2): 179 - 181. [Full Text] [PDF]
|
|
|
|
 |
|
 J. P. Stains and R. Civitelli Gap Junctions Regulate Extracellular Signal-regulated Kinase Signaling to Affect Gene Transcription Mol. Biol. Cell, January 1, 2005; 16(1): 64 - 72. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
B. N.G Giepmans Gap junctions and connexin-interacting proteins Cardiovasc Res, May 1, 2004; 62(2): 233 - 245. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S. Dhein Pharmacology of gap junctions in the cardiovascular system Cardiovasc Res, May 1, 2004; 62(2): 287 - 298. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
R. Schulz and G. Heusch Connexin 43 and ischemic preconditioning Cardiovasc Res, May 1, 2004; 62(2): 335 - 344. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
D. Garcia-Dorado, A. Rodriguez-Sinovas, and M. Ruiz-Meana Gap junction-mediated spread of cell injury and death during myocardial ischemia-reperfusion Cardiovasc Res, February 15, 2004; 61(3): 386 - 401. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 B. W. Doble, X. Dang, P. Ping, R. R. Fandrich, B. E. Nickel, Y. Jin, P. A. Cattini, and E. Kardami Phosphorylation of serine 262 in the gap junction protein connexin-43 regulates DNA synthesis in cell-cell contact forming cardiomyocytes J. Cell Sci., January 22, 2004; 117(3): 507 - 514. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
T. Miura, Y. Ohnuma, A. Kuno, M. Tanno, Y. Ichikawa, Y. Nakamura, T. Yano, T. Miki, J. Sakamoto, and K. Shimamoto Protective role of gap junctions in preconditioning against myocardial infarction Am J Physiol Heart Circ Physiol, January 1, 2004; 286(1): H214 - H221. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J. C. SAEZ, V. M. BERTHOUD, M. C. BRANES, A. D. MARTINEZ, and E. C. BEYER Plasma Membrane Channels Formed by Connexins: Their Regulation and Functions Physiol Rev, October 1, 2003; 83(4): 1359 - 1400. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
K. A. Detillieux, F. Sheikh, E. Kardami, and P. A. Cattini Biological activities of fibroblast growth factor-2 in the adult myocardium Cardiovasc Res, January 1, 2003; 57(1): 8 - 19. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
U. Schwanke, I. Konietzka, A. Duschin, X. Li, R. Schulz, and G. Heusch No ischemic preconditioning in heterozygous connexin43-deficient mice Am J Physiol Heart Circ Physiol, October 1, 2002; 283 (4): H1740 - H1742. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
D. Garcia-Dorado, M. Ruiz-Meana, F. Padilla, A. Rodriguez-Sinovas, and M. Mirabet Gap junction-mediated intercellular communication in ischemic preconditioning Cardiovasc Res, August 15, 2002; 55(3): 456 - 465. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 C. P. Baines, J. Zhang, G.-W. Wang, Y.-T. Zheng, J. X. Xiu, E. M. Cardwell, R. Bolli, and P. Ping Mitochondrial PKC{epsilon} and MAPK Form Signaling Modules in the Murine Heart: Enhanced Mitochondrial PKC{epsilon}-MAPK Interactions and Differential MAPK Activation in PKC{epsilon}-Induced Cardioprotection Circ. Res., March 8, 2002; 90(4): 390 - 397. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J. L. Tuttle and J. C. Falcone Nitric oxide release during {alpha}1-adrenoceptor-mediated constriction of arterioles Am J Physiol Heart Circ Physiol, August 1, 2001; 281(2): H873 - H881. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
M. Ruiz-Meana, D. Garcia-Dorado, S. Lane, P. Pina, J. Inserte, M. Mirabet, and J. Soler-Soler Persistence of gap junction communication during myocardial ischemia Am J Physiol Heart Circ Physiol, June 1, 2001; 280(6): H2563 - H2571. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
R. M. Fryer, Y. Wang, A. K. Hsu, and G. J. Gross Essential activation of PKC-{delta} in opioid-initiated cardioprotection Am J Physiol Heart Circ Physiol, March 1, 2001; 280(3): H1346 - H1353. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J. B. Strait III, J. L. Martin, A. Bayer, R. Mestril, D. M. Eble, and A. M. Samarel Role of protein kinase C-{epsilon} in hypertrophy of cultured neonatal rat ventricular myocytes Am J Physiol Heart Circ Physiol, February 1, 2001; 280(2): H756 - H766. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
Z.-S. Jiang, R. R. Padua, H. Ju, B. W. Doble, Y. Jin, J. Hao, P. A. Cattini, I. M. C. Dixon, and E. Kardami Acute protection of ischemic heart by FGF-2: involvement of FGF-2 receptors and protein kinase C Am J Physiol Heart Circ Physiol, March 1, 2002; 282(3): H1071 - H1080. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
T. M. Vondriska, J. Zhang, C. Song, X.-L. Tang, X. Cao, C. P. Baines, J. M. Pass, S. Wang, R. Bolli, and P. Ping Protein Kinase C {epsilon}-Src Modules Direct Signal Transduction in Nitric Oxide-Induced Cardioprotection : Complex Formation as a Means for Cardioprotective Signaling Circ. Res., June 22, 2001; 88(12): 1306 - 1313. [Abstract] [Full Text] [PDF]
|
|