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Recognition of Double-stranded RNA by Human Toll-like Receptor 3 and Downstream Receptor Signaling Requires Multimerization and an Acidic pH

de Bouteiller, Odette, Merck, Estelle, Hasan, Uzma A., Hubac, Sylvain, Benguigui, Barbara, Trinchieri, Giorgio, Bates, Elizabeth E.M., Caux, Christophe·2005·Journal of Biological Chemistry
DOI 10.1074/jbc.m507163200
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Abstract

Studies involving Toll-like receptor 3 (TLR3)-deficient mice suggest that this receptor binds double-stranded RNA. In the present study, we analyzed ligand/receptor interactions and receptor-proximal events leading to TLR3 activation. The mutagenesis approach showed that certain cysteine residues and glycosylation in TLR3 amino-terminal leucine-rich repeats were necessary for ligand-induced signaling. Furthermore, inactive mutants had a dominant negative effect, suggesting that the signaling module is a multimer. We constructed a chimeric molecule fusing the amino-terminal ectodomain of TLR3 to the transmembrane and carboxyl terminal domains of CD32a containing an immunoreceptor tyrosine-based motif. Expression of TLR3-CD32 in HEK293T cells and the myeloid cell line U937 resulted in surface localization of the receptor, whereas the nonrecombinant molecule was intracellularly localized. The synthetic double-stranded RNAs poly(I-C) and poly(A-U) induced calcium mobilization in a TLR3-CD32 stably transfected U937 clone but not in control cells transfected with other constructs. An anti-TLR3 antibody also induced Ca(2+) flux but only when cross-linked by a secondary anti-immunoglobulin antibody, confirming that multimerization by the ligand is a requirement for signaling. The inhibitors of lysosome maturation, bafilomycin and chloroquine, inhibited the poly(I-C)-induced biological response in immune cells, showing that TLR3 interacted with its ligand in acidic subcellular compart…

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  • In phagocytic cells, deacidification of endosomes using chloroquine reduces endosomal toll-like receptor response.

    “In phagocytic cells, the deacidification of endosomes using pharmacological inhibitors like chloroquine has been shown to reduce endosomal toll-like receptor (TLR) response (de Bouteiller et al., 2005; Fox, 2019; Kuznik et al., 2011; Wozniacka et al., 2006; Yang et al., 2016)”

    From NHE6 depletion corrects ApoE4-mediated synaptic impairments and reduces amyloid plaque load (2021) · cites this paper as de Bouteiller et al., 2005

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Metadata

Authors
de Bouteiller, O., Merck, E., Hasan, U. A., Hubac, S., Benguigui, B., Trinchieri, G., Bates, E. E. M., & Caux, C.
Venue
Journal of Biological Chemistry
Publisher
Elsevier BV
Published
2005-11
Volume
280
Issue
46
Pages
38133-38145
ISSN
0021-9258
DOI
10.1074/jbc.m507163200
Cited by
220
References
6
Language
en
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