Skip to main content
SpringerLink
Log in

A novel role of G protein-coupled receptor kinase 5 in urotensin II-stimulated cellular hypertrophy in H9c2UT cells

  • Published:
Molecular and Cellular Biochemistry Aims and scope Submit manuscript

Abstract

Urotensin II (UII) is a neural hormone that induces cardiac hypertrophy and may be involved in the pathogenesis of cardiac remodeling and heart failure. Hypertrophy has been linked to histone deacetylase 5 (HDAC5) phosphorylation and nuclear factor κB (NF-κB) translocation, both of which are predominantly mediated by G protein-coupled receptor kinase 5 (GRK5). In the present study, we found that UII rapidly and strongly stimulated nuclear export of HDAC5 and nuclear import of NF-κB in H9c2 cells overexpressing the urotensin II receptor (H9c2UT). Hence, we hypothesized that GRK5 and its signaling pathway may play a role in UII-mediated cellular hypertrophy. H9c2UT cells were transduced with a GRK5 small hairpin RNA interference recombinant lentivirus, resulting in the down-regulation of GRK5. Under UII stimulation, reduced levels of GRK5 in H9c2UT cells led to suppression of UII-mediated HDAC5 phosphorylation and activation of the NF-κB signaling pathway. In contrast, UII-mediated activations of ERK1/2 and GSK3α/β were not affected by down-regulation of GRK5. In a cellular hypertrophy assay, down-regulation of GRK5 significantly suppressed UII-mediated hypertrophy of H9c2UT cells. Furthermore, UII-mediated cellular hypertrophy was inhibited by amlexanox, a selective GRK5 inhibitor, in H9c2UT cells and neonatal cardiomyocytes. Our results suggest that GRK5 may be involved in a UII-mediated hypertrophic response via activation of NF-κB and HDAC5 at least in part by ERK1/2 and GSK3α/β-independent pathways.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6

Similar content being viewed by others

References

  1. Douglas SA, Naselsky D, Ao Z, Disa J, Herold CL, Lynch F, Aiyar NV (2004) Identification and pharmacological characterization of native, functional human urotensin-II receptors in rhabdomyosarcoma cell lines. Br J Pharmacol 142:921–932

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  2. Ong KL, Lam KS, Cheung BM (2005) Urotensin II: its function in health and its role in disease. Cardiovasc Drugs Ther 19:65–75

    Article  CAS  PubMed  Google Scholar 

  3. Tzanidis A, Hannan RD, Thomas WG, Onan D, Autelitano DJ, See F, Kelly DJ, Gilbert RE, Krum H (2003) Direct actions of urotensin II on the heart: implications for cardiac fibrosis and hypertrophy. Circ Res 93:246–253

    Article  CAS  PubMed  Google Scholar 

  4. Onan D, Pipolo L, Yang E, Hannan RD, Thomas WG (2004) Urotensin II promotes hypertrophy of cardiac myocytes via mitogen-activated protein kinases. Mol Endocrinol 18:2344–2354

    Article  CAS  PubMed  Google Scholar 

  5. Gruson D, Ginion A, Decroly N, Lause P, Vanoverschelde JL, Ketelslegers JM, Bertrand L, Thissen JP (2010) Urotensin II induction of adult cardiomyocytes hypertrophy involves the Akt/GSK-3beta signaling pathway. Peptides 31:1326–1333

    Article  CAS  PubMed  Google Scholar 

  6. Ping P, Anzai T, Gao M, Hammond HK (1997) Adenylyl cyclase and G protein receptor kinase expression during development of heart failure. Am J Physiol 273:H707–H717

    CAS  PubMed  Google Scholar 

  7. Rockman HA, Choi DJ, Rahman NU, Akhter SA, Lefkowitz RJ, Koch WJ (1996) Receptor-specific in vivo desensitization by the G protein-coupled receptor kinase-5 in transgenic mice. Proc Natl Acad Sci USA 93:9954–9959

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  8. Eckhart AD, Duncan SJ, Penn RB, Benovic JL, Lefkowitz RJ, Koch WJ (2000) Hybrid transgenic mice reveal in vivo specificity of G protein-coupled receptor kinases in the heart. Circ Res 86:43–50

    Article  CAS  PubMed  Google Scholar 

  9. Parameswaran N, Pao CS, Leonhard KS, Kang DS, Kratz M, Ley SC, Benovic JL (2006) Arrestin-2 and G protein-coupled receptor kinase 5 interact with NFkappaB1 p105 and negatively regulate lipopolysaccharide-stimulated ERK1/2 activation in macrophages. J Biol Chem 281:34159–34170

    Article  CAS  PubMed  Google Scholar 

  10. Martini JS, Raake P, Vinge LE, DeGeorge BR Jr, Chuprun JK, Harris DM (2008) Uncovering G protein-coupled receptor kinase-5 as a histone deacetylase kinase in the nucleus of cardiomyocytes. Proc Natl Acad Sci USA 105:12457–12462

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  11. Sorriento D, Ciccarelli M, Santulli G, Campanile A, Altobelli GG, Cimini V, Galasso G, Astone D, Piscione F, Pastore L, Trimarco B, Iaccarino G (2008) The G-protein-coupled receptor kinase 5 inhibits NFkappaB transcriptional activity by inducing nuclear accumulation of IkappaB alpha. Proc Natl Acad Sci USA 105:17818–17823

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  12. Barthet G, Carrat G, Cassier E, Barker B, Gaven F, Pillot M, Framery B, Pellissier LP, Augier J, Kang DS, Claeysen S, Reiter E, Banères JL, Benovic JL, Marin P, Bockaert J, Dumuis A (2009) Beta-arrestin1 phosphorylation by GRK5 regulates G protein-independent 5-HT4 receptor signaling. EMBO J 28:2706–2718

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  13. Patial S, Luo J, Porter KJ, Benovic JL, Parameswaran N (2009) G-protein-coupled-receptor kinases mediate TNFα-induced NFκB signalling via direct interaction with and phosphorylation of IκBα. Biochem J 425:169–178

    Article  PubMed  PubMed Central  Google Scholar 

  14. Chen X, Zhu H, Yuan M, Fu J, Zhou Y, Ma L (2010) G-protein-coupled receptor kinase 5 phosphorylates p53 and inhibits DNA damage-induced apoptosis. J Biol Chem 285:12823–12830

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  15. Michal AM, So CH, Beeharry N, Shankar H, Mashayekhi R, Yen TJ, Benovic JL (2012) G Protein-coupled receptor kinase 5 is localized to centrosomes and regulates cell cycle progression. J Biol Chem 287:6928–6940

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  16. Burkhalter MD, Fralish GB, Premont RT, Caron MG, Philipp M (2013) Grk5l controls heart development by limiting mTOR signaling during symmetry breaking. Cell Rep 4:625–632

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  17. Traynham CJ, Hullmann J, Koch WJ (2016) Canonical and non-canonical actions of GRK5 in the heart. J Mol Cell Cardiol 92:196–202

    Article  CAS  PubMed  Google Scholar 

  18. Oh KS, Lee JH, Yi KY, Lim CJ, Lee S, Park CH, Seo HW, Lee BH (2015) The orally active urotensin receptor antagonist, KR36676, attenuates cellular and cardiac hypertrophy. Br J Pharmacol 172:2618–2633

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  19. Kim J, Ahn S, Ren XR, Whalen EJ, Reiter E, Wei H, Lefkowitz RJ (2005) Functional antagonism of different G protein-coupled receptor kinases for beta-arrestin-mediated angiotensin II receptor signaling. Proc Natl Acad Sci USA 102:1442–1447

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  20. Ren XR, Reiter E, Ahn S, Kim J, Chen W, Lefkowitz RJ (2005) Different G protein-coupled receptor kinases govern G protein and beta-arrestin-mediated signaling of V2 vasopressin receptor. Proc Natl Acad Sci USA 102:1448–1453

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  21. Shenoy SK, Drake MT, Nelson CD, Houtz DA, Xiao K, Madabushi S, Reiter E, Premont RT, Lichtarge O, Lefkowitz RJ (2006) Beta-arrestin-dependent, G protein-independent ERK1/2 activation by the beta2 adrenergic receptor. J Biol Chem 281:1261–1273

    Article  CAS  PubMed  Google Scholar 

  22. Zhang CL, McKinsey TA, Chang S, Antos CL, Hill JA, Olson EN (2002) Class II histone deacetylases act as signal-responsive repressors of cardiac hypertrophy. Cell 110:479–488

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  23. McKinsey TA, Olson EN (2004) Dual roles of histone deacetylases in the control of cardiac growth. Novartis Found Symp 259:132–141

    Article  CAS  PubMed  Google Scholar 

  24. Backs J, Song K, Bezprozvannaya S, Chang S, Olson EN (2006) Cam kinase II selectively signals to histone deacetylase 4 during cardiomyocyte hypertrophy. J Clin Investig 116:1853–1864

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  25. Olson EN, Backs J, McKinsey TA (2006) Control of cardiac hypertrophy and heart failure by histone acetylation/deacetylation. Novartis Found Symp 274:3–12

    Article  CAS  PubMed  Google Scholar 

  26. Zhu W, Tilley DG, Myers VD, Coleman RC, Feldman AM (2013) Arginine vasopressin enhances cell survival via a G protein-coupled receptor kinase 2/β-arrestin1/extracellular-regulated kinase 1/2-dependent pathway in H9c2 cells. Mol Pharmacol 84:227–235

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  27. Purcell NH, Tang G, Yu C, Mercurio F, DiDonato JA, Lin A (2001) Activation of NF-kappa B is required for hypertrophic growth of primary rat neonatal ventricular cardiomyocytes. Proc Natl Acad Sci USA 98:6668–6673

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  28. Islam KN, Bae JW, Gao E, Koch WJ (2013) Regulation of nuclear factor κB (NF-κB) in the nucleus of cardiomyocytes by G protein-coupled receptor kinase 5 (GRK5). J Biol Chem 288:35683–35689

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  29. Homan KT, Wu E, Cannavo A, Koch WJ, Tesmer JJ (2014) Identification and characterization of amlexanox as a G protein-coupled receptor kinase 5 inhibitor. Molecules 19:16937–16949

    Article  PubMed  PubMed Central  Google Scholar 

  30. Shi H, Han Q, Xu J, Liu W, Chu T, Zhao L (2016) Urotensin II induction of neonatal cardiomyocyte hypertrophy involves the CaMKII/PLN/SERCA 2a signaling pathway. Gene 583:8–14

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgments

This study was supported by the Bio & Medical Technology Development Program (2014-M3A9A9073788) of the National Research Foundation (NRF) funded by Korean government, Republic of Korea.

Author information

Authors and Affiliations

  1. Center for Drug Discovery Technology, Korea Research Institute of Chemical Technology, Daejeon, Republic of Korea

    Cheon Ho Park, Ju Hee Lee, Mi Young Lee, Jeong Hyun Lee, Byung Ho Lee & Kwang-Seok Oh

  2. Graduate School of New Drug Discovery and Development, Chungnam National University, Daejeon, Republic of Korea

    Byung Ho Lee

  3. Department of Medicinal and Pharmaceutical Chemistry, University of Science and Technology, Daejeon, Republic of Korea

    Kwang-Seok Oh

Authors
  1. Cheon Ho Park
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Ju Hee Lee
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Mi Young Lee
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Jeong Hyun Lee
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Byung Ho Lee
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Kwang-Seok Oh
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding authors

Correspondence to Byung Ho Lee or Kwang-Seok Oh.

Ethics declarations

Conflict of interest

The authors declare that they have no conflict of interest.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Park, C.H., Lee, J.H., Lee, M.Y. et al. A novel role of G protein-coupled receptor kinase 5 in urotensin II-stimulated cellular hypertrophy in H9c2UT cells. Mol Cell Biochem 422, 151–160 (2016). https://doi.org/10.1007/s11010-016-2814-y

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11010-016-2814-y

Keywords

Search

Navigation