Structural basis for TIR domain–mediated innate immune signaling by Toll-like receptor adaptors TRIF and TRAM

M Mohammad K. Manik (Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School) M Mengqi Pan (School of Chemistry and Molecular Biosciences, University of Queensland) L Le Xiao (Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School) W Weixi Gu (School of Chemistry and Molecular Biosciences, University of Queensland) H Hyoyoung Kim (School of Chemistry and Molecular Biosciences, University of Queensland) S Sabrina Pospich (Department of Structural Biochemistry, Max Planck Institute of Molecular Physiology) A Andrew Hedger (School of Chemistry and Molecular Biosciences, University of Queensland) P Parimala R. Vajjhala (School of Chemistry and Molecular Biosciences, University of Queensland) M Morris Y. L. Lee (School of Chemistry and Molecular Biosciences, University of Queensland) X Xiaoqi Qian (School of Chemistry and Molecular Biosciences, University of Queensland) M Michael J. Landsberg (School of Chemistry and Molecular Biosciences, University of Queensland) T Thomas Ve (Institute for Biomedicine and Glycomics, Griffith University) J Jeffrey D. Nanson (School of Chemistry and Molecular Biosciences, University of Queensland) S Stefan Raunser K Katryn J. Stacey (School of Chemistry and Molecular Biosciences, University of Queensland) H Hao Wu B Bostjan Kobe (The University of Queensland, School of Chemistry and Molecular Biosciences, Australian Infectious Diseases Research Centre and Institute for Molecular Bioscience)

Abstract

Innate immunity relies on Toll-like receptors (TLRs) to detect pathogen-associated molecular patterns. The TIR (Toll/interleukin-1 receptor) domain-containing TLR adaptors TRIF (TIR domain–containing adaptor-inducing interferon-β) and TRAM (TRIF-related adaptor molecule) are essential for MyD88-independent TLR signaling. However, the structural basis of TRIF and TRAM TIR domain–based signaling remains unclear. Here, we present cryo-EM structures of filaments formed by TRIF and TRAM TIR domains at resolutions of 3.3 Å and 5.6 Å, respectively. Both structures reveal two-stranded parallel helical arrangements. Functional studies underscore the importance of intrastrand interactions, mediated by the BB-loop, and interstrand interactions in TLR4-mediated signaling. We also report the crystal structure of the monomeric TRAM TIR domain bearing the BB loop mutation C117H, which reveals conformational differences consistent with its inactivity. Our findings suggest a unified signaling mechanism by the TIR domains of the four signaling TLR adaptors MyD88, MAL, TRIF, and TRAM and reveal potential therapeutic targets for immunity-related disorders.

Article Details

Volume / Issue Vol. 122, Issue 2
Published January 14, 2025
ISSN 0027-8424
Publisher National Academy of Sciences

Authors (17)

M

Mohammad K. Manik

Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School

M

Mengqi Pan

School of Chemistry and Molecular Biosciences, University of Queensland

L

Le Xiao

Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School

W

Weixi Gu

School of Chemistry and Molecular Biosciences, University of Queensland

H

Hyoyoung Kim

School of Chemistry and Molecular Biosciences, University of Queensland

S

Sabrina Pospich

Department of Structural Biochemistry, Max Planck Institute of Molecular Physiology

A

Andrew Hedger

School of Chemistry and Molecular Biosciences, University of Queensland

P

Parimala R. Vajjhala

School of Chemistry and Molecular Biosciences, University of Queensland

M

Morris Y. L. Lee

School of Chemistry and Molecular Biosciences, University of Queensland

X

Xiaoqi Qian

School of Chemistry and Molecular Biosciences, University of Queensland

M

Michael J. Landsberg

School of Chemistry and Molecular Biosciences, University of Queensland

T

Thomas Ve

Institute for Biomedicine and Glycomics, Griffith University

J

Jeffrey D. Nanson

School of Chemistry and Molecular Biosciences, University of Queensland

S

Stefan Raunser

K

Katryn J. Stacey

School of Chemistry and Molecular Biosciences, University of Queensland

H

Hao Wu

B

Bostjan Kobe

The University of Queensland, School of Chemistry and Molecular Biosciences, Australian Infectious Diseases Research Centre and Institute for Molecular Bioscience