TLR Lecture.ppt
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Transcript of TLR Lecture.ppt
Toll-Like Receptors (TLR)
Toll-Like Receptor Signaling• Toll receptor initially discovered in Drosophila as
important receptor in dorso-ventral embryonic pattern– Toll mutants refers to the fact that these mutants could not
establish a proper dorsal-ventral axis– Toll in German means ‘great’, apparently this was one of the
words describing the scientists’ enthusiasm after observing the mutant flies
• Hoffman and colleagues showed that Toll-mutant flies susceptible to fungal infections
• Mammalian homologues discovered and designated as Toll-Like Receptors (TLR)
• TLRs recognize specific patterns in pathogens which are not observed in mammals
Toll-Like Receptors
Toll-Like Receptors
• Large molecules found in outer membrane of Gram- negative bacteria
• Comprised of a lipid and saccharide component• Highly immunogenic• Recognized by TLR4• Can cause septic shock and lead to death• Often referred to as Endotoxin since it is not
secreted but is a byproduct of bacterial lysis• Great variability among different bacterial strains
LipoPolySaccharides (LPS)
• TLR3, TLR7 and TLR8 detect viral nucleic acids
• Found in intracellular membranes since viral nucleic acids are endogenously generated
• Note the TIR domains of these TLRs face the cytosol of the cell
• Poly I:C is an agonist for TLR3 that mimics binding of dsRNA to TLR3
Viral Nucleic Acids
Cytoplasmic tails of TLRs show similarities to IL-1 receptor (TIR)
Common adaptor to TLRs is MyD88
Crucial proline residue in all TLR TIR domains, except TLR3
If substituted with histidine, no signaling occurs
All TLRs likely have a MyD88 pathway (TLR3 is an exception)
TLR4 has a MyD88 independent pathway as well
CytoplasmicTIR domain
TLRs and TIR Domain
MyD88 knockout mice have no response to LPS
MyD88 is essential to all inflammatory signaling pathways
MyD88s, a splice variant of MyD88 down regulates the inflammatory response
MyD88 interacts with TIR domain of TLR and recruits IRAK-4, IRAK-1 and TRAF-6
MyD88 Adaptor
• 4 IRAKs known, IRAK1, IRAK-2, IRAK-M and IRAK-4
• IRAK are serine/threonine kinases• IRAK-4 phosphorylates IRAK-1• IRAK-M plays an inhibitory role in TLR
signaling • IRAK-4 phosphorylates IRAK-1• TRAF6 is a member of the TNF receptor
associated factor (TRAF) family• TRAF6 interacts with IRAK-1 and gets
activated• Release of TRAF6/IRAK-1 ensues
IRAK and TRAF6
• TRAF6/IRAK-1 complex associates with 3 proteins
– TAK1 (TGF-B activated kinase)
– TAB1 (TAK1 binding proteins)
– TAB2 (TAK1 binding proteins)
• Large complex associates with membrane
• Eventually IRAK-1 stays in membrane while TRAF6/TAK1/TAB1/TAB2 move to cytosol
• E2 Ligases such as Ubc13 and Uev1A join further enlarging complex
IRAK and TRAF6 Release
TAK-1 Activation• The enlarged complex that
includes
– TRAF6, TAK1, TAB1, TAB2, Ubc13, Uev1A activate TAK1
• Activated TAK1 phosphorylates IKK complex
• Activated TAK1 can also phosphorylate MAP Kinases
• IKK complex consists of IKK, and /NEMO
• IB phosphorylation results in NF- B translocation to nucleus
MyD88 Independent Pathway MyD88 Knock out mice do not produce inflammatory cytokines such as TNF-
However with TLR4 stimulation NF-B and JNK delayed activity occurs
This strongly suggests the existence of 2 pathways in TLR signaling
a MyD88 dependent pathway (early)
a MyD88 independent pathway (late)
TLR3 stimulation also exchibits a MyD88 independent pathway
TLR4
• Figures obtained from:
• Takeda and Akira, 2004
• Wikipedia
• Kuby, 6th edition
Acknowledgements