nri0604-poster.

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Toll-like receptor signalling

Luke A. J. O’Neill
Toll-like receptors (TLRs) recognize microbial products during
infection and initiate signalling pathways that culminate in the
increased expression of immune and inflammatory genes. TLRs that
recognize nucleic acids signal from endosomes, whereas cell-sur-
face TLRs sense lipids and proteins. Two major signalling pathways
have been detailed. The core pathway activated by most TLRs leads
to activation of the transcription factor nuclear factor-κB (NF-κB) and
the mitogen-activated protein (MAP) kinases p38 and JNK. These sig-
nalling cascades increase the expression of many pro-inflammatory
genes. The second pathway is activated by TLR3 and TLR4 and leads
to activation of both NF-κB and another transcription factor interferon
regulatory factor 3 (IRF3), allowing for an additional set of genes to be
induced, including antiviral genes such as interferon-β (IFN-β ). In this
way, TLRs can tailor the innate response to pathogens. updated september 2005

TLR ligands Adaptor molecules specify

Host mRNA HSP60 (?), fibrinogen (?), β-defensin (?) mRNA (?) TLR-signalling effects
and hyaluronic-acid fragments (?)
Parasites GPI-linked proteins GPI-linked proteins Toxoplasma gondii profilin Central to TLR signalling are the receptor-
Fungi Zymosan Zymosan proximal adaptors, MyD88, MAL (also known
as TIRAP), TRIF (also known as TICAM1) and
Viruses Double-stranded RNA F protein Single-stranded RNA Herpes-virus DNA TRAM (also known as TICAM2 or TIRP). A fifth
Bacteria Lipopolysaccharides Triacyl lipopeptides Diacyl lipopeptides Flagellin Uropathogenic bacteria CpG DNA sequences adaptor, SARM, has been described, but at
Small molecules Imiquimod and Resiquimod least in the case of nematodes, it does not
Other Synthetic CpG oligodeoxynucleotides seem to be recruited by TLRs. MyD88 is
widely used by TLRs, although MyD88
RP105 TLR6 signalling initiated by TLR2 and TLR4, but not
CD14 TLR4 TLR1 TLR2 TLR2 TLR6 TLR2 CD36
other TLRs, also involves the adaptor MAL.
TLR5 TLR11 MyD88 recruits IRAKs, ultimately leading to
MD2 SIGIRR ST2
the activation of MAP3 kinases, two of which
have been identified, MEKK3 and TAK1, which
Cell membrane activate NF-κB and the MAP kinases p38 and
Endosome

RAC

RAC
RAC
BTK
TRAM

MAL

MAL

MAL
MAL
TRIF

MyD88

MyD88
JNK. Activation of TAK1 by IRAKs requires

MyD88

MyD88
MyD88

MyD88
TLR3 PI3K PI3K PI3K TLR7 or TLR8 TRAF6, as well as the ubiquitylation of both
TRAF6 and TAK1. BTK and PI3K also

RIP2

RIP2

RIP2
participate in TLR signalling, although their
RIP2

precise role has yet to be defined. The kinase

MyD88s RIP2 has been shown to be essential for
TRIAD3A signalling by certain TLRs, in particular TLR2
and TLR4. By contrast, RIP1 is involved only
MyD88s
TRIF

in TLR3 signalling. The adaptor molecule TRIF

BTK
MyD88
RIP3
ROS IRAK4 engages the protein kinase TBK1, leading to
IRF3 activation downstream of TLR3 and TLR4

IRAK-M
signalling, and can also interact directly with
TRAF6. Interestingly, TRIF-mediated signalling
downstream of TLR4 also requires the adaptor
TRAM, whereas TLR3 signalling through TRIF
RIP1 TRAF6 PI3K TBK1 TRAF6 TRAF6 does not. Another IRF, IRF8, has recently been
IRAK1 IRAK2 MyD88s shown to be activated by TLR9 through MyD88.
TOLLIP
TLR9 The process of NF-κB activation downstream
of TLR4 ligation has added complexity, with
the early response involving MyD88 and MAL,
AKT and the later response involving TRAM and
UEV1A TRAF6 UBC13 A20 TRAF6
TRIF. Although much has been learnt about
IRAK1
the importance of adaptors in defining unique
ECSIT innate immune responses, the full
consequences of specific adaptor usage by

MyD88
IKK1 TAB2 TLRs have yet to be resolved.
NEMO

? IRF3 IRF5 ASK1

TAB1

IKK2 TAK1 MEKK3

Negative regulation of
TRAF6 ? TLR signalling
TRIAD3A
TLR signalling is negatively regulated by
IKK1 various proteins. The cell-surface receptors
NEMO

MKK3 MKK7 IRF5 IRF8 ST2 (also known as T1) and SIGIRR function
IKK2 as inhibitory receptors, sequestering proteins
IκB from signalling complexes and preventing
IRF7 TLR2, TLR4 and TLR9 signalling. IRAK-M,
p50 p65 NF-κB TOLLIP and a splice variant of MyD88, known
IκB
as MyD88s, probably interfere with the
p38 p50 p65 NF-κB JNK
recruitment and activation of IRAK4 and
IRAK1. Recently, TRIAD3A, a RING-finger
E3 ligase, has been shown to promote
ubiquitylation of TLR4 and TLR9, targeting
these TLRs for degradation and thereby
negatively regulating the intensity and duration
IL-6 TNF IL-6
IL-12p40 p50 p65 COX2 IL-12p40 IFN-α of TLR signalling. The balance between
p50 p65 IRF3 IFN-β IRF5 TNF IL-18 IRF5 TNF IRF7 activation and inhibition is likely to be the
IFN-β
NF-κB- key determinant of signal strength.
NF-κB-
binding motif Nuclear membrane
binding motif ISRE ISRE ISRE ISRE

Late phase Early phase

For the purpose of clarity, the pathways depicted here are limited to those that are well understood and known to have an
important role in TLR signalling. For a more comprehensive overview of the TLR-signalling pathways, please see the
review article by Shizuo Akira and Kiyoshi Takeda in the July issue of Nature Reviews Immunology. This article, as well
as the poster, is freely available online for 3 months as part of a focus on TLR signalling: www.nature.com/nri/focus.tlr.
In addition, the online version of the poster will be updated every 3 months to include new data that become available.
Designed by Neil Smith. Edited by Karen Honey and Lucy Bird. © 2004 Nature Publishing Group.
ASK1, apoptosis signal-regulating kinase 1; BTK, Bruton’s tyrosine kinase; COX2, cyclo-oxygenase 2; ECSIT, evolutionarily conserved signalling intermediate in Toll pathways; GPI, glycosylphosphatidylinositol;
HSP60, heat-shock protein 60; IFN, interferon; IκB, inhibitor of NF-κB; IKK, inhibitor of NF-κB kinase; IRAK, IL-1-receptor-associated kinase; IL, interleukin; IRF, IFN regulatory factor; JNK, JUN N-terminal kinase;
MAL, MyD88 adaptor-like protein; MAP, mitogen-activated protein; MEKK3, MAPK/ERKK kinase 3; MKK, MAP kinase kinase; MyD88, myeloid differentiation primary-response gene 88; NEMO, NF-κB essential modulator;
NF-κB, nuclear factor-κB; PI3K, phosphatidylinositol 3-kinase; RIP, receptor-interacting protein; ROS, reactive oxygen species; SARM, sterile α and armadillo motifs; SIGIRR, single immunoglobulin IL-1-receptor-related
molecule; TAB, TAK1-binding protein; TAK1, TGF-β-activated protein kinase 1; TANK, TRAF-family-member-associated NF-κB activator; TBK1, TANK-binding kinase 1; TGF-β, transforming growth factor-β; TICAM, TIR-
domain-containing adaptor molecule; TIR, Toll/IL-1 receptor; TIRAP, TIR-domain-containing adaptor protein; TLR, Toll-like receptor; TNF, tumour-necrosis factor; TOLLIP, Toll-interacting protein; TRAF, TNF-receptor-
associated factor; TRAM, TRIF-related adaptor molecule; TRIF, TIR-domain-containing adaptor protein inducing IFN-β; UBC13, ubiquitin-conjugating enzyme 13; UEV1A, ubiquitin-conjugating enzyme E2 variant 1.