Modulation of Nod2-dependent NF-B signaling by the actin cytoskeleton

Sylvie Legrand-Poels^1,*, Gaelle Kustermans¹, Françoise Bex², Elisabeth Kremmer³, Thomas A. Kufer⁴, and Jacques Piette¹

¹ Laboratory of Virology and Immunology, CBIG-GIGA, University of Liège, Liège, Belgium
² Laboratory of Microbiology, Institute for Microbiological Research J.-M. Wiame, University of Brussels, Brussels, Belgium
³ GSF-Institut für Molekulare Immunologie, Marchioninistr. 25, 81377 München, Germany
⁴ Unité de Pathogénie Microbienne Moléculaire, Institut Pasteur and INSERM, U786, Paris 75724 CEDEX 15, France

^* Author for correspondence (e-mail: S.Legrand{at}ulg.ac.be)

Abstract: Actin disruption by CytochalasinD (CytD) and LatrunculinB (LatB) induced NF-B activation in myelomonocytic and intestinal epithelial cells. In an attempt to elucidate the mechanism by which actin disruption induced IKK activation, we studied the human Nod2 protein, which was able to induce NF-B activation and whose expression was restricted to myelomonocytic and intestinal epithelial cells. Nod2 is thought to play key roles in pathogen defence through sensing bacteria and generating an inflammatory immune response. We showed that actin disruption by CytD significantly and specifically increased Nod2-mediated NF-B signaling. Nod2 was fully partitioned in the Triton-X-100-insoluble fraction but translocated into the soluble fraction after CytD treatment, demonstrating that the presence of Nod2 in the detergent-insoluble pellet was specific to actin cytoskeleton. Confocal analysis also revealed a Nod2 colocalization with membrane-associated F-actin. Colocalization and co-immunoprecipitation assays with endogenous Rac1 have shown that Nod2 associated with activated Rac1 in membrane ruffles through both its N-terminal caspase recruitment domains (CARD) and C-terminal leucine-rich repeats (LRRs). Membrane ruffle disruption by a Rac1 dominant negative form primed Nod2-dependent NF-B signaling. The recruitment of Nod2 in Rac-induced dynamic cytoskeletal structures could be a strategy to both repress the Nod2-dependent NF-B signaling in unstimulated cells and rapidly mobilize Nod2 during bacterial infection.

Key Words: Actin cytoskeleton • NF-B • Nod2 • Rac1

The intermediate filament protein, vimentin, is a regulator of NOD2 activity.

C. Stevens, P. Henderson, E. R. Nimmo, D. C. Soares, B. Dogan, K. W. Simpson, J. C. Barrett, International Inflammatory Bowel Disease Genetics, D. C. Wilson, and J. Satsangi (2013)
Gut 62, 695-707
 |  Abstract »  |  Full Text »  |  PDF »

Disruption of asymmetric dimethylarginine-induced RelA/P65 association with actin in endothelial cells.

W. Guo, D. Zhang, L. Wang, Y. Zhang, and W. Liu (2013)
Acta Biochim Biophys Sin 45, 229-235
 |  Abstract »  |  Full Text »  |  PDF »

Mutational analysis of human NOD1 and NOD2 NACHT domains reveals different modes of activation.

B. Zurek, M. Proell, R. N. Wagner, R. Schwarzenbacher, and T. A. Kufer (2012)
Innate Immunity 18, 100-111
 |  Abstract »  |  Full Text »  |  PDF »

Retinoic Acid-induced Gene-I (RIG-I) Associates with Nucleotide-binding Oligomerization Domain-2 (NOD2) to Negatively Regulate Inflammatory Signaling.

S. A. Morosky, J. Zhu, A. Mukherjee, S. N. Sarkar, and C. B. Coyne (2011)
J. Biol. Chem. 286, 28574-28583
 |  Abstract »  |  Full Text »  |  PDF »

CIN85 drives B cell responses by linking BCR signals to the canonical NF-{kappa}B pathway.

K. Kometani, T. Yamada, Y. Sasaki, T. Yokosuka, T. Saito, K. Rajewsky, M. Ishiai, M. Hikida, and T. Kurosaki (2011)
J. Exp. Med. 208, 1447-1457
 |  Abstract »  |  Full Text »  |  PDF »

Pattern-recognition Receptors in Pulp Defense.

M.- J. Staquet, F. Carrouel, J.- F. Keller, C. Baudouin, P. Msika, F. Bleicher, T. A. Kufer, and J.- C. Farges (2011)
Advances in Dental Research 23, 296-301
 |  Abstract »  |  Full Text »  |  PDF »

Expression of NOD2 is increased in inflamed human dental pulps and lipoteichoic acid-stimulated odontoblast-like cells.

J.-F. Keller, F. Carrouel, M.-J. Staquet, T. A. Kufer, C. Baudouin, P. Msika, F. Bleicher, and J.-C. Farges (2011)
Innate Immunity 17, 29-34
 |  Abstract »  |  PDF »

DUOX2-derived reactive oxygen species are effectors of NOD2-mediated antibacterial responses.

S. Lipinski, A. Till, C. Sina, A. Arlt, H. Grasberger, S. Schreiber, and P. Rosenstiel (2009)
J. Cell Sci. 122, 3522-3530
 |  Abstract »  |  Full Text »  |  PDF »

Critical Role of NOD2 in Regulating the Immune Response to Staphylococcus aureus.

H. S. Deshmukh, J. B. Hamburger, S. H. Ahn, D. G. McCafferty, S. R. Yang, and V. G. Fowler Jr. (2009)
Infect. Immun. 77, 1376-1382
 |  Abstract »  |  Full Text »  |  PDF »

Retinoic Acid-induced Gene-1 (RIG-I) Associates with the Actin Cytoskeleton via Caspase Activation and Recruitment Domain-dependent Interactions.

A. Mukherjee, S. A. Morosky, L. Shen, C. R. Weber, J. R. Turner, K. S. Kim, T. Wang, and C. B. Coyne (2009)
J. Biol. Chem. 284, 6486-6494
 |  Abstract »  |  Full Text »  |  PDF »

Role of actin cytoskeleton in LPS-induced NF-{kappa}B activation and nitric oxide production in murine macrophages.

S. M. Eswarappa, V. Pareek, and D. Chakravortty (2008)
Innate Immunity 14, 309-318
 |  Abstract »  |  PDF »

{beta}-PIX and Rac1 GTPase Mediate Trafficking and Negative Regulation of NOD2.

J. Eitel, M. Krull, A. C. Hocke, P. D. N'Guessan, J. Zahlten, B. Schmeck, H. Slevogt, S. Hippenstiel, N. Suttorp, and B. Opitz (2008)
J. Immunol. 181, 2664-2671
 |  Abstract »  |  Full Text »  |  PDF »

A role for membrane-bound CD147 in NOD2-mediated recognition of bacterial cytoinvasion.

A. Till, P. Rosenstiel, K. Brautigam, C. Sina, G. Jacobs, H.-H. Oberg, D. Seegert, T. Chakraborty, and S. Schreiber (2008)
J. Cell Sci. 121, 487-495
 |  Abstract »  |  Full Text »  |  PDF »