Sensing bacterial flagellin by membrane and soluble orthologs of toll-like receptor 5 in rainbow trout (Onchorhynchus mikiss)

Tadayuki Tsujita, Hironobu Tsukada, Miki Nakao, Hiroyuki Oshiumi, Misako Matsumoto, Tsukasa Seya

Research output: Contribution to journalArticle

147 Citations (Scopus)

Abstract

Rainbow trout (Onchorhynchus mikiss) possess two genes encoding putative leucine-rich repeat (LRR)-containing proteins similar to human TLR5. Molecular cloning of these two LRR proteins suggested the presence of a TLR5-like membrane form (rtTLR5M) and a soluble form (rtTLR5S). Here we elucidated the primary structures and the unique combinational functions of these fish versions of TLR5. The LRR regions of rtTLR5S and rtTLR5M exhibited 81% homology and relatively high (35.6 and 33.7%) homology to the extracellular domains of human TLR5 (huTLR5). Thus, two distinct genes encode the TLR5 orthologs in fish, one of which has a consensus intracellular domain (TIR). In order to test their functions, we constructed fusion proteins with the LRR region of rtTLR5S (S-chimera) or thai of rtTLR5M and the TIR of huTLR5 (M-chimera). The S- and M-chimeras expressed in HeLa or CHO cells signaled the presence of Vibrio anguillarum flagellin, resulting in NF-κB activation. rtTLR5M was ubiquitously expressed, whereas rtTLR5S was predominantly expressed in the liver. In the hepatoma cell lines of the rainbow trout RTH-149, stimulation of rtTLR5M with V. anguillarum or its flagellin allowed the up-regulation of rtTLR5S. Flagellin-mediated NF-κB activation was more significant in the presence of or simultaneous expression of rtTLR5S. Therefore, a two-step flagellin response occurred for host defense against bacterial infection in fish: (a) flagellin first induced basal activation of NF-κB via membrane TLR5, facilitating the production of soluble TLR5 and minimal acute phase proteins, and (b) the inducible soluble TLR5 amplifies membrane TLR5-mediated cellular responses in a positive feedback fashion.

Original languageEnglish
Pages (from-to)48588-48597
Number of pages10
JournalJournal of Biological Chemistry
Volume279
Issue number47
DOIs
Publication statusPublished - Nov 19 2004

Fingerprint

Toll-Like Receptor 5
Flagellin
Oncorhynchus mykiss
Membranes
Leucine
Fish
Fishes
Chemical activation
Gene encoding
Step response
Vibrio
CHO Cells
Acute-Phase Proteins
Cloning
Molecular Cloning
HeLa Cells
Bacterial Infections
Liver
Genes
Hepatocellular Carcinoma

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Sensing bacterial flagellin by membrane and soluble orthologs of toll-like receptor 5 in rainbow trout (Onchorhynchus mikiss). / Tsujita, Tadayuki; Tsukada, Hironobu; Nakao, Miki; Oshiumi, Hiroyuki; Matsumoto, Misako; Seya, Tsukasa.

In: Journal of Biological Chemistry, Vol. 279, No. 47, 19.11.2004, p. 48588-48597.

Research output: Contribution to journalArticle

Tsujita, Tadayuki ; Tsukada, Hironobu ; Nakao, Miki ; Oshiumi, Hiroyuki ; Matsumoto, Misako ; Seya, Tsukasa. / Sensing bacterial flagellin by membrane and soluble orthologs of toll-like receptor 5 in rainbow trout (Onchorhynchus mikiss). In: Journal of Biological Chemistry. 2004 ; Vol. 279, No. 47. pp. 48588-48597.
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