Note to users. If you're seeing this message, it means that your browser cannot find this page's style/presentation instructions -- or possibly that you are using a browser that does not support current Web standards. Find out more about why this message is appearing, and what you can do to make your experience of our site the best it can be.
Subscribe

Logo for

J. Cell Biol. 177 (2): 265-275

Copyright © 2007 by the Rockefeller University Press.

Article

The interaction between the ER membrane protein UNC93B and TLR3, 7, and 9 is crucial for TLR signaling

Melanie M. Brinkmann1, Eric Spooner1, Kasper Hoebe2, Bruce Beutler2, Hidde L. Ploegh1, , and You-Me Kim1

1 Whitehead Institute for Biomedical Research, Cambridge, MA 02142
2 Department of Immunology, The Scripps Research Institute, La Jolla, CA 92037

Correspondence to Hidde L. Ploegh: ploegh{at}wi.mit.edu; or You-Me Kim: ykim{at}wi.mit.edu

Abstract: Toll-like receptors (TLRs) sense the presence of microbial and viral pathogens by signal transduction mechanisms that remain to be fully elucidated. A single point mutation (H412R) in the polytopic endoplasmic reticulum (ER)–resident membrane protein UNC93B abolishes signaling via TLR3, 7, and 9. We show that UNC93B specifically interacts with TLR3, 7, 9, and 13, whereas introduction of the point mutation H412R in UNC93B abolishes their interactions. We establish the physical interaction of the intracellular TLRs with UNC93B in splenocytes and bone marrow–derived dendritic cells. Further, by expressing chimeric TLRs, we show that TLR3 and 9 bind to UNC93B via their transmembrane domains. We propose that a physical association between UNC93B and TLRs in the ER is essential for proper TLR signaling.

Abbreviations used in this paper: ATCC, American Type Culture Collection; BM-DC, bone marrow–derived dendritic cell; HEK, human embryonic kidney; HIFS, heat-inactivated fetal calf serum; LPS, lipopolysaccharide; MHC, major histocompatibility complex; MS, mass spectrometry; TEV, Tobacco Etch virus; TIR, Toll-interleukin 1 receptor; TLR, Toll-like receptor.

THIS ARTICLE HAS BEEN CITED BY OTHER ARTICLES:
Acidic Amino Acid Residues in the Juxtamembrane Region of the Nucleotide-Sensing TLRs Are Important for UNC93B1 Binding and Signaling.
J. Kim, J. Huh, M. Hwang, E.-H. Kwon, D.-J. Jung, M. M. Brinkmann, M. H. Jang, H. L. Ploegh, and Y.-M. Kim (2013)
J. Immunol. 190, 5287-5295
   Abstract »    Full Text »    PDF »
Interferon-{gamma} and Granulocyte/Monocyte Colony-stimulating Factor Production by Natural Killer Cells Involves Different Signaling Pathways and the Adaptor Stimulator of Interferon Genes (STING).
F. Souza-Fonseca-Guimaraes, M. Parlato, R. B. de Oliveira, D. Golenbock, K. Fitzgerald, I. N. Shalova, S. K. Biswas, J.-M. Cavaillon, and M. Adib-Conquy (2013)
J. Biol. Chem. 288, 10715-10721
   Abstract »    Full Text »    PDF »
Recognition of pathogen-associated nucleic acids by endosomal nucleic acid-sensing toll-like receptors.
X. He, H. Jia, Z. Jing, and D. Liu (2013)
Acta Biochim Biophys Sin 45, 241-258
   Abstract »    Full Text »    PDF »
TLR9 mediates cellular protection by modulating energy metabolism in cardiomyocytes and neurons.
Y. Shintani, A. Kapoor, M. Kaneko, R. T. Smolenski, F. D'Acquisto, S. R. Coppen, N. Harada-Shoji, H. J. Lee, C. Thiemermann, S. Takashima, et al. (2013)
PNAS 110, 5109-5114
   Abstract »    Full Text »    PDF »
Essential role for Toll-like receptor 7 (TLR7)-unique cysteines in an intramolecular disulfide bond, proteolytic cleavage and RNA sensing.
A. Kanno, C. Yamamoto, M. Onji, R. Fukui, S.-i. Saitoh, Y. Motoi, T. Shibata, F. Matsumoto, T. Muta, and K. Miyake (2013)
Int. Immunol.
   Abstract »    Full Text »    PDF »
Cell-Specific TLR9 Trafficking in Primary APCs of Transgenic TLR9-GFP Mice.
A. M. Avalos, O. Kirak, J. M. Oelkers, M. C. Pils, Y.-M. Kim, M. Ottinger, R. Jaenisch, H. L. Ploegh, and M. M. Brinkmann (2013)
J. Immunol. 190, 695-702
   Abstract »    Full Text »    PDF »
The Role of UNC93B1 Protein in Surface Localization of TLR3 Receptor and in Cell Priming to Nucleic Acid Agonists.
J. Pohar, N. Pirher, M. Bencina, M. Mancek-Keber, and R. Jerala (2013)
J. Biol. Chem. 288, 442-454
   Abstract »    Full Text »    PDF »
UNC93B1 mediates differential trafficking of endosomal TLRs.
B. L. Lee, J. E. Moon, J. H. Shu, L. Yuan, Z. R. Newman, R. Schekman, and G. M. Barton (2013)
eLife Sci 2, e00291
   Abstract »    Full Text »    PDF »
IgM+IgD+CD27+ B cells are markedly reduced in IRAK-4-, MyD88-, and TIRAP- but not UNC-93B-deficient patients.
S. Weller, M. Bonnet, H. Delagreverie, L. Israel, M. Chrabieh, L. Marodi, C. Rodriguez-Gallego, B.-Z. Garty, C. Roifman, A. C. Issekutz, et al. (2012)
Blood 120, 4992-5001
   Abstract »    Full Text »    PDF »
Sequence specific detection of bacterial 23S ribosomal RNA by TLR13.
X.-D. Li and Z. J. Chen (2012)
eLife Sci 1, e00102
   Abstract »    Full Text »    PDF »
Failure To Induce IFN-{beta} Production during Staphylococcus aureus Infection Contributes to Pathogenicity.
A. Kaplan, J. Ma, P. Kyme, A. J. Wolf, C. A. Becker, C. W. Tseng, G. Y. Liu, and D. M. Underhill (2012)
J. Immunol. 189, 4537-4545
   Abstract »    Full Text »    PDF »
Proteolytic Processing Regulates Toll-like Receptor 3 Stability and Endosomal Localization.
R. Qi, D. Singh, and C. C. Kao (2012)
J. Biol. Chem. 287, 32617-32629
   Abstract »    Full Text »    PDF »
Cutting Edge: TLR13 Is a Receptor for Bacterial RNA.
A. Hidmark, A. von Saint Paul, and A. H. Dalpke (2012)
J. Immunol. 189, 2717-2721
   Abstract »    Full Text »    PDF »
TLR13 Recognizes Bacterial 23S rRNA Devoid of Erythromycin Resistance-Forming Modification.
M. Oldenburg, A. Kruger, R. Ferstl, A. Kaufmann, G. Nees, A. Sigmund, B. Bathke, H. Lauterbach, M. Suter, S. Dreher, et al. (2012)
Science 337, 1111-1115
   Abstract »    Full Text »    PDF »
Cleavage of Toll-like receptor 3 by cathepsins B and H is essential for signaling.
A. Garcia-Cattaneo, F.-X. Gobert, M. Muller, F. Toscano, M. Flores, A. Lescure, E. Del Nery, and P. Benaroch (2012)
PNAS 109, 9053-9058
   Abstract »    Full Text »    PDF »
Role of Specific Innate Immune Responses in Herpes Simplex Virus Infection of the Central Nervous System.
J. P. Wang, G. N. Bowen, S. Zhou, A. Cerny, A. Zacharia, D. M. Knipe, R. W. Finberg, and E. A. Kurt-Jones (2012)
J. Virol. 86, 2273-2281
   Abstract »    Full Text »    PDF »
The role of DNA sensing and innate immune receptor TLR9 in otitis media.
A. Leichtle, M. Hernandez, J. Lee, K. Pak, N. J. Webster, B. Wollenberg, S. I. Wasserman, and A. F. Ryan (2012)
Innate Immunity 18, 3-13
   Abstract »    Full Text »    PDF »
Requirement of UNC93B1 Reveals a Critical Role for TLR7 in Host Resistance to Primary Infection with Trypanosoma cruzi.
B. C. Caetano, B. B. Carmo, M. B. Melo, A. Cerny, S. L. dos Santos, D. C. Bartholomeu, D. T. Golenbock, and R. T. Gazzinelli (2011)
J. Immunol. 187, 1903-1911
   Abstract »    Full Text »    PDF »
BAD-LAMP is a novel biomarker of nonactivated human plasmacytoid dendritic cells.
A. Defays, A. David, A. de Gassart, F. De Angelis Rigotti, T. Wenger, V. Camossetto, P. Brousset, T. Petrella, M. Dalod, E. Gatti, et al. (2011)
Blood 118, 609-617
   Abstract »    Full Text »    PDF »
Toll-Like Receptors Participate in Macrophage Activation and Intracellular Control of Leishmania (Viannia) panamensis.
C. Gallego, D. Golenbock, M. A. Gomez, and N. G. Saravia (2011)
Infect. Immun. 79, 2871-2879
   Abstract »    Full Text »    PDF »
Small molecule Toll-like receptor 7 agonists localize to the MHC class II loading compartment of human plasmacytoid dendritic cells.
C. Russo, I. Cornella-Taracido, L. Galli-Stampino, R. Jain, E. Harrington, Y. Isome, S. Tavarini, C. Sammicheli, S. Nuti, M. L. Mbow, et al. (2011)
Blood 117, 5683-5691
   Abstract »    Full Text »    PDF »
Nucleic acid recognition by Toll-like receptors is coupled to stepwise processing by cathepsins and asparagine endopeptidase.
S. E. Ewald, A. Engel, J. Lee, M. Wang, M. Bogyo, and G. M. Barton (2011)
J. Exp. Med. 208, 643-651
   Abstract »    Full Text »    PDF »
A Point Mutation in the Amino Terminus of TLR7 Abolishes Signaling without Affecting Ligand Binding.
C. Iavarone, K. Ramsauer, A. V. Kubarenko, J. C. Debasitis, I. Leykin, A. N. R. Weber, O. M. Siggs, B. Beutler, P. Zhang, G. Otten, et al. (2011)
J. Immunol. 186, 4213-4222
   Abstract »    Full Text »    PDF »
Optimal oligonucleotide sequences for TLR9 inhibitory activity in human cells: lack of correlation with TLR9 binding.
R. F. Ashman, J. A. Goeken, E. Latz, and P. Lenert (2011)
Int. Immunol. 23, 203-214
   Abstract »    Full Text »    PDF »
Therapeutical targeting of nucleic acid-sensing Toll-like receptors prevents experimental cerebral malaria.
B. S. Franklin, S. T. Ishizaka, M. Lamphier, F. Gusovsky, H. Hansen, J. Rose, W. Zheng, M. A. Ataide, R. B. de Oliveira, D. T. Golenbock, et al. (2011)
PNAS 108, 3689-3694
   Abstract »    Full Text »    PDF »
A Novel Toll-like Receptor That Recognizes Vesicular Stomatitis Virus.
Z. Shi, Z. Cai, A. Sanchez, T. Zhang, S. Wen, J. Wang, J. Yang, S. Fu, and D. Zhang (2011)
J. Biol. Chem. 286, 4517-4524
   Abstract »    Full Text »    PDF »
UNC93B1 Is Essential for TLR11 Activation and IL-12-dependent Host Resistance to Toxoplasma gondii.
R. Pifer, A. Benson, C. R. Sturge, and F. Yarovinsky (2011)
J. Biol. Chem. 286, 3307-3314
   Abstract »    Full Text »    PDF »
Bortezomib suppresses function and survival of plasmacytoid dendritic cells by targeting intracellular trafficking of Toll-like receptors and endoplasmic reticulum homeostasis.
M. Hirai, N. Kadowaki, T. Kitawaki, H. Fujita, A. Takaori-Kondo, R. Fukui, K. Miyake, T. Maeda, S. Kamihira, Y. Miyachi, et al. (2011)
Blood 117, 500-509
   Abstract »    Full Text »    PDF »
CD14 is a coreceptor of Toll-like receptors 7 and 9.
C. L. Baumann, I. M. Aspalter, O. Sharif, A. Pichlmair, S. Bluml, F. Grebien, M. Bruckner, P. Pasierbek, K. Aumayr, M. Planyavsky, et al. (2010)
J. Exp. Med. 207, 2689-2701
   Abstract »    Full Text »    PDF »
Secretion of the Human Toll-like Receptor 3 Ectodomain Is Affected by Single Nucleotide Polymorphisms and Regulated by Unc93b1.
R. Qi, S. Hoose, J. Schreiter, K. V. Sawant, R. Lamb, C. T. Ranjith-Kumar, J. Mills, L. S. Mateo, J. L. Jordan, and C. C. Kao (2010)
J. Biol. Chem. 285, 36635-36644
   Abstract »    Full Text »    PDF »
Bifurcation of Toll-Like Receptor 9 Signaling by Adaptor Protein 3.
M. Sasai, M. M. Linehan, and A. Iwasaki (2010)
Science 329, 1530-1534
   Abstract »    Full Text »    PDF »
Chicken TLR21 Is an Innate CpG DNA Receptor Distinct from Mammalian TLR9.
A. M. Keestra, M. R. de Zoete, L. I. Bouwman, and J. P. M. van Putten (2010)
J. Immunol. 185, 460-467
   Abstract »    Full Text »    PDF »
Up-regulation of the endoplasmic reticulum transmembrane protein UNC93B in the B cells of patients with active systemic lupus erythematosus.
S. Nakano, S. Morimoto, S. Suzuki, T. Watanabe, H. Amano, and Y. Takasaki (2010)
Rheumatology 49, 876-881
   Abstract »    Full Text »    PDF »
Cotrafficking of SV2 and Synaptotagmin at the Synapse.
J. Yao, A. Nowack, P. Kensel-Hammes, R. G. Gardner, and S. M. Bajjalieh (2010)
J. Neurosci. 30, 5569-5578
   Abstract »    Full Text »    PDF »
Murine B Cell Response to TLR7 Ligands Depends on an IFN-{beta} Feedback Loop.
N. M. Green, A. Laws, K. Kiefer, L. Busconi, Y.-M. Kim, M. M. Brinkmann, E. H. Trail, K. Yasuda, S. R. Christensen, M. J. Shlomchik, et al. (2009)
J. Immunol. 183, 1569-1576
   Abstract »    Full Text »    PDF »
Unc93B1 biases Toll-like receptor responses to nucleic acid in dendritic cells toward DNA- but against RNA-sensing.
R. Fukui, S.-i. Saitoh, F. Matsumoto, H. Kozuka-Hata, M. Oyama, K. Tabeta, B. Beutler, and K. Miyake (2009)
J. Exp. Med. 206, 1339-1350
   Abstract »    Full Text »    PDF »
Bacterial DNA Activates Endothelial Cells and Promotes Neutrophil Adherence through TLR9 Signaling.
D. El Kebir, L. Jozsef, W. Pan, L. Wang, and J. G. Filep (2009)
J. Immunol. 182, 4386-4394
   Abstract »    Full Text »    PDF »
The roles of TLRs, RLRs and NLRs in pathogen recognition.
T. Kawai and S. Akira (2009)
Int. Immunol. 21, 317-337
   Abstract »    Full Text »    PDF »
A single base mutation in the PRAT4A gene reveals differential interaction of PRAT4A with Toll-like receptors.
T. Kiyokawa, S. Akashi-Takamura, T. Shibata, F. Matsumoto, C. Nishitani, Y. Kuroki, Y. Seto, and K. Miyake (2008)
Int. Immunol. 20, 1407-1415
   Abstract »    Full Text »    PDF »
Mouse cytomegalovirus inhibits beta interferon (IFN-{beta}) gene expression and controls activation pathways of the IFN-{beta} enhanceosome.
V. T. K. Le, M. Trilling, A. Zimmermann, and H. Hengel (2008)
J. Gen. Virol. 89, 1131-1141
   Abstract »    Full Text »    PDF »
The interaction between the ER membrane protein UNC93B and TLR3, 7, and 9 is crucial for TLR signaling.
M. M. Brinkmann, E. Spooner, K. Hoebe, B. Beutler, H. L. Ploegh, and Y.-M. Kim (2007)
J. Exp. Med. 204, i14
   Full Text »

To Advertise     Find Products

Science Signaling. ISSN 1937-9145 (online), 1945-0877 (print). Pre-2008: Science's STKE. ISSN 1525-8882