Identification of an N-terminal recognition site in TLR9 that contributes to CpG-DNA-mediated receptor activation.
Although it is well established that TLR9 recognizes CpG-DNA, the structural details of ligand-receptor interaction are still mostly unknown. The extracellular domain of TLR9 is composed of 25 leucine-rich repeat (LRR) motifs, 5 of which bear inserting sequences that do not conform to the LRR consensus motif. In this study, we show that the functional integrity of the extracellular domain of murine TLR9 is lost by deletion of individual LRR motifs. When deleting only the inserting sequences, we observed that LRR2, 5, and 8 contribute to receptor activation by CpG-DNA. The latter deletions did not affect receptor dimerization but inhibited CpG-DNA binding. On the basis of a homology modeling approach, we furthermore identify a positively charged region in the N terminus that is essential for CpG-DNA-induced TLR9 activation. This interaction site mirrors findings previously shown for the structural recognition of dsRNA by TLR3 and hints toward a general principle of nucleic acid recognition by the respective TLR.
Peter
Mirjam E
ME
Department of Medical Microbiology and Hygiene, University Heidelberg, Heidelberg, Germany.
Kubarenko
Andriy V
AV
Weber
Alexander N R
AN
Dalpke
Alexander H
AH
D000595:Amino Acid Sequence; D000818:Animals; D002460:Cell Line; D017124:Conserved Sequence; D018899:CpG Islands / Q000235:genetics*; D004247:DNA / Q000235:genetics*; D017353:Gene Deletion; D006801:Humans; D007930:Leucine / Q000235:genetics / Q000378:metabolism; D051379:Mice; D008958:Models, Molecular; D008969:Molecular Sequence Data; D009154:Mutation / Q000235:genetics; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D016415:Sequence Alignment; D040681:Structural Homology, Protein; D051196:Toll-Like Receptor 3 / Q000737:chemistry / Q000378:metabolism; D051217:Toll-Like Receptor 9 / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*
C495370:Tlr9 protein, mouse; D051196:Toll-Like Receptor 3; D051217:Toll-Like Receptor 9; D004247:DNA; D007930:Leucine