Subscribe to RSS
Endothelial CSN5 impairs NF-κB activation and monocyte adhesion to endothelial cells and is highly expressed in human atherosclerotic lesionsFinancial support: This work was supported by the Deutsche Forschungsgemeinschaft (DFG) International graduate school grant IRTG1508/1-TP6 to J.B. and Y.A., by the Alexander von Humboldt Foundation to H.N., and by the Netherlands Organization for Health Research and Development to M.W. M.W. is an established investigator of the Netherlands Heart Foundation.
20 February 2013
Accepted after minor revision: 05 April 2013
30 November 2017 (online)
The COP9 signalosome (CSN), a multifunctional protein complex involved in the regulation of cullin-RING-E3 ubiquitin ligases (CRLs), has emerged as a regulator of NF-κB signalling. As NF-κB drives the expression of pro-inflammatory and pro-atherosclerotic genes, we probed the yet unknown role of the CSN, in particular CSN5, on NF-KB-mediated atherogenic responses in endothelial cells. Co-immunoprecipitation in human umbilical vein endothelial cells (HUVECs) revealed the presence of a super-complex between IKK and CSN, which dissociates upon TNF-α stimulation. Furthermore, CSN5 silencing enhanced TNF-α-induced IKB-α degradation and NF-κB activity in luci-ferase reporter assays. This was paralleled by an increased NF-KB-driven upregulation of atherogenic chemokines and adhesion molecules, as measured by qPCR and flow cytometry, and translated into an enhanced arrest of THP-1 monocytes on TNF-α-stimulated, CSN5-depleted HUVECs. Reverse effects on NF-κB activity and THP-1 arrest were seen upon CSN5 overexpression. Finally, double-immunostaining confirmed the expression of CSN subunits in the endothelium of human atherosclerotic lesions, and revealed an increased expression of CSN5 which correlated with atheroprogression. In conclusion, endothelial CSN5 attenuates NF-KB-dependent pro-inflammatory gene expression and monocyte arrest on stimulated endothelial cells in vitro, suggesting that CSN5 might serve as a negative regulator of atherogenesis.
Note: The review process for this manuscript was fully handled by G. Y. H. Lip, Editor in Chief.
§ These authors share senior-authorship and contributed equally to this work.
* Current address: Department of Medical Biochemistry, Academic Medical Center, University of Amsterdam, Amsterdam, the Netherlands.
- 1 Hansson GK, Hermansson A. The immune system in atherosclerosis. Nat Immunol 2011; 12: 204-212.
- 2 Weber C, Noels H. Atherosclerosis: current pathogenesis and therapeutic options. Nat Med 2011; 17: 1410-1422.
- 3 Bernhagen J, Krohn R, Lue H. et al. MIF is a noncognate ligand of CXC chemokine receptors in inflammatory and atherogenic cell recruitment. Nat Med 2007; 13: 587-596.
- 4 de Winther MP, Kanters E, Kraal G. et al. Nuclear factor kappaB signalling in atherogenesis. Arterioscler Thromb Vasc Biol 2005; 25: 904-914.
- 5 Bonizzi G, Karin M. The two NF-kappaB activation pathways and their role in innate and adaptive immunity. Trends Immunol 2004; 25: 280-288.
- 6 Karin M, Ben-Neriah Y. Phosphorylation meets ubiquitination: the control of NF-[kappa]B activity. Annu Rev Immunol 2000; 18: 621-663.
- 7 Sun SC, Ganchi PA, Ballard DW. et al. NF-kappa B controls expression of inhibitor I kappa B alpha: evidence for an inducible autoregulatory pathway. Science 1993; 259: 1912-1915.
- 8 Wei N, Serino G, Deng XW. The COP9 signalosome: more than a protease. Trends Biochem Sci 2008; 33: 592-600.
- 9 Schweitzer K, Bozko PM, Dubiel W. et al. CSN controls NF-kappaB by deubiquitinylation of IkappaBalpha. Embo J 2007; 26: 1532-1541.
- 10 Kapelari B, Bech-Otschir D, Hegerl R. et al. Electron microscopy and subunit-subunit interaction studies reveal a first architecture of COP9 signalosome. J Mol Biol 2000; 300: 1169-1178.
- 11 Wei N, Deng XW. COP9: a new genetic locus involved in light-regulated development and gene expression in arabidopsis. Plant Cell 1992; 04: 1507-1518.
- 12 Lyapina S, Cope G, Shevchenko A. et al. Promotion of NEDD-CUL1 conjugate cleavage by COP9 signalosome. Science 2001; 292: 1382-1385.
- 13 Cope GA, Suh GS, Aravind L. et al. Role of predicted metalloprotease motif of Jab1/Csn5 in cleavage of Nedd8 from Cul1. Science 2002; 298: 608-611.
- 14 Bosu DR, Kipreos ET. Cullin-RING ubiquitin ligases: global regulation and activation cycles. Cell Div 2008; 03: 7.
- 15 Carrano AC, Eytan E, Hershko A. et al. SKP2 is required for ubiquitin-mediated degradation of the CDK inhibitor p27. Nat Cell Biol 1999; 01: 193-199.
- 16 Read MA, Brownell JE, Gladysheva TB. et al. Nedd8 modification of cul-1 activates SCF(beta(TrCP))-dependent ubiquitination of IkappaBalpha. Mol Cell Biol 2000; 20: 2326-2333.
- 17 Wu K, Chen A, Pan ZQ. Conjugation of Nedd8 to CUL1 enhances the ability of the ROC1-CUL1 complex to promote ubiquitin polymerisation. J Biol Chem 2000; 275: 32317-32324.
- 18 Podust VN, Brownell JE, Gladysheva TB. et al. A Nedd8 conjugation pathway is essential for proteolytic targeting of p27Kip1 by ubiquitination. Proc Natl Acad Sci USA 2000; 97: 4579-4584.
- 19 Zhou C, Seibert V, Geyer R. et al. The fission yeast COP9/signalosome is involved in cullin modification by ubiquitin-related Ned8p. BMC Biochem 2001; 02: 7.
- 20 Yang X, Menon S, Lykke-Andersen K. et al. The COP9 signalosome inhibits p27(kip1) degradation and impedes G1-S phase progression via deneddylation of SCF Cul1. Curr Biol 2002; 12: 667-672.
- 21 Wee S, Geyer RK, Toda T. et al. CSN facilitates Cullin-RING ubiquitin ligase function by counteracting autocatalytic adapter instability. Nat Cell Biol 2005; 07: 387-391.
- 22 Wu JT, Lin HC, Hu YC. et al. Neddylation and deneddylation regulate Cul1 and Cul3 protein accumulation. Nat Cell Biol 2005; 07: 1014-1020.
- 23 Denti S, Fernandez-Sanchez ME, Rogge L. et al. The COP9 signalosome regulates Skp2 levels and proliferation of human cells. J Biol Chem 2006; 281: 32188-32196.
- 24 Schmidt MW, McQuary PR, Wee S. et al. F-box-directed CRL complex assembly and regulation by the CSN and CAND1. Mol Cell 2009; 35: 586-597.
- 25 Emberley ED, Mosadeghi R, Deshaies RJ. Deconjugation of Nedd8 from Cul1 is directly regulated by Skp1-F-box and substrate, and the COP9 signalosome inhibits deneddylated SCF by a noncatalytic mechanism. J Biol Chem 2012; 287: 29679-29689.
- 26 Peth A, Berndt C, Henke W. et al. Downregulation of COP9 signalosome subunits differentially affects the CSN complex and target protein stability. BMC Biochem 2007; 08: 27.
- 27 Zhou C, Wee S, Rhee E. et al. Fission yeast COP9/signalosome suppresses cullin activity through recruitment of the deubiquitylating enzyme Ubp12p. Mol Cell 2003; 11: 927-938.
- 28 Groisman R, Polanowska J, Kuraoka I. et al. The ubiquitin ligase activity in the DDB2 and CSA complexes is differentially regulated by the COP9 signalosome in response to DNA damage. Cell 2003; 113: 357-367.
- 29 Hetfeld BK, Helfrich A, Kapelari B. et al. The zinc finger of the CSN-associated deubiquitinating enzyme USP15 is essential to rescue the E3 ligase Rbx1. Curr Biol 2005; 15: 1217-1221.
- 30 Kim BC, Lee HJ, Park SH. et al. Jab1/CSN5, a component of the COP9 signalosome, regulates transforming growth factor beta signalling by binding to Smad7 and promoting its degradation. Mol Cell Biol 2004; 24: 2251-2262.
- 31 Adler AS, Littlepage LE, Lin M. et al. CSN5 isopeptidase activity links COP9 signalosome activation to breast cancer progression. Cancer Res 2008; 68: 506-515.
- 32 Panattoni M, Sanvito F, Basso V. et al. Targeted inactivation of the COP9 signalosome impairs multiple stages of T cell development. J Exp Med 2008; 205: 465-477.
- 33 Gu L, Okada Y, Clinton SK. et al. Absence of monocyte chemoattractant protein-1 reduces atherosclerosis in low density lipoprotein receptor-deficient mice. Mol Cell 1998; 02: 275-281.
- 34 Aiello RJ, Bourassa PA, Lindsey S. et al. Monocyte chemoattractant protein-1 accelerates atherosclerosis in apolipoprotein E-deficient mice. Arterioscler Thromb Vasc Biol 1999; 19: 1518-1525.
- 35 Johnson RC, Chapman SM, Dong ZM. et al. Absence of P-selectin delays fatty streak formation in mice. J Clin Invest 1997; 99: 1037-1043.
- 36 Collins RG, Velji R, Guevara NV. et al. P-Selectin or intercellular adhesion molecule (ICAM)-1 deficiency substantially protects against atherosclerosis in apolipoprotein E-deficient mice. J Exp Med 2000; 191: 189-194.
- 37 Cybulsky MI, Iiyama K, Li H. et al. A major role for VCAM-1, but not ICAM-1, in early atherosclerosis. J Clin Invest 2001; 107: 1255-1262.
- 38 Oron E, Mannervik M, Rencus S. et al. COP9 signalosome subunits 4 and 5 regulate multiple pleiotropic pathways in Drosophila melanogaster. Development 2002; 129: 4399-4409.
- 39 Peth A, Boettcher JP, Dubiel W. Ubiquitin-dependent proteolysis of the microtubule end-binding protein 1, EB1, is controlled by the COP9 signalosome: possible consequences for microtubule filament stability. J Mol Biol 2007; 368: 550-563.
- 40 Burger-Kentischer A, Finkelmeier D, Thiele M. et al. Binding of JAB1/CSN5 to MIF is mediated by the MPN domain but is independent of the JAMM motif. FEBS Lett 2005; 579: 1693-1701.
- 41 Tomoda K, Kato JY, Tatsumi E. et al. The Jab1/COP9 signalosome subcomplex is a downstream mediator of Bcr-Abl kinase activity and facilitates cell-cycle progression. Blood 2005; 105: 775-783.
- 42 Tomoda K, Kubota Y, Arata Y. et al. The cytoplasmic shuttling and subsequent degradation of p27Kip1 mediated by Jab1/CSN5 and the COP9 signalosome complex. J Biol Chem 2002; 277: 2302-2310.
- 43 Orel L, Neumeier H, Hochrainer K. et al. Crosstalk between the NF-kappaB activating IKK-complex and the CSN signalosome. J Cell Mol Med 2010; 14: 1555-1568.
- 44 Burger-Kentischer A, Goebel H, Seiler R. et al. Expression of macrophage migration inhibitory factor in different stages of human atherosclerosis. Circulation 2002; 105: 1561-1566.
- 45 Gareus R, Kotsaki E, Xanthoulea S. et al. Endothelial cell-specific NF-kappaB inhibition protects mice from atherosclerosis. Cell Metab 2008; 08: 372-383.
- 46 Kanters E, Pasparakis M, Gijbels MJ. et al. Inhibition of NF-kappaB activation in macrophages increases atherosclerosis in LDL receptor-deficient mice. J Clin Invest 2003; 112: 1176-1185.
- 47 Claret FX, Hibi M, Dhut S. et al. A new group of conserved coactivators that increase the specificity of AP-1 transcription factors. Nature 1996; 383: 453-457.
- 48 Adler AS, Lin M, Horlings H. et al. Genetic regulators of large-scale transcriptional signatures in cancer. Nat Genet 2006; 38: 421-430.
- 49 Kleemann R, Hausser A, Geiger G. et al. Intracellular action of the cytokine MIF to modulate AP-1 activity and the cell cycle through Jab1. Nature 2000; 408: 211-216.
- 50 Noels H, Bernhagen J, Weber C. Macrophage migration inhibitory factor: a noncanonical chemokine important in atherosclerosis. Trends Cardiovasc Med 2009; 19: 76-86.
- 51 Lue H, Thiele M, Franz J. et al. Macrophage migration inhibitory factor (MIF) promotes cell survival by activation of the Akt pathway and role for CSN5/JAB1 in the control of autocrine MIF activity. Oncogene 2007; 26: 5046-5059.