Innate immunity provides a first line of defense against pathogen- or danger-associated molecular patterns. Among known innate receptors, Toll-like receptors (TLRs) and NOD-like receptors (NLRs) share a common structural motif that consists of multiple leucine-rich repeats (LRRs) with some variability in amino- acid residues and length of each repeat. Structural studies of LRRs of variable lymphocyte receptors (VLRs) in jawless fish have also shown that LRR domains are important in ligand recognition. Thus, the LRRs of TLRs and NLRs are essential for interaction with agonist ligands in order to initiate cellular signaling in innate immunity. Furthermore, when the LRR domain is disengaged from the signaling domains, it can negatively regulate innate immunity, by competitively binding to ligands. Thus, the ultimate goal of this study is to advance understanding of the structure and function of LRRs in TLRs and NLRs in ligand-induced activation and signaling. Structural and biophysical characterization of LRR domains in TLR1/2/3/4/5/6 have been accomplished by several groups, including our own, but major unresolved questions remain concerning how TLR7/8/9 interact with and are regulated by nucleic acid and other ligands in response to microbial pathogens. Compared to TLRs, structural investigation of NLRs has been extremely limited. To date, only one NLR LRR structure, that of NLRX1, has been solved and we have also shown that it directly binds RNA ligands. With our extensive expertise in expression, purification, crystallization and characterization of diverse LRR proteins, we will investigate structural, biophysical and functional properties of LRR domains in TLR7/8/9 and NLRC3/4/5, as well as advance our already productive studies of TLR5 and NLRX1. Such findings will address the structural and functional mechanisms that arise from the specific interactions of TLR7/8/9 and NLRC3/4/5 with their cognate ligands and lead to understanding of receptor activation and regulation in innate immunity. We will capitalize on our previous studies of TLR5 and NLRX1 to answer key questions about their signaling mechanisms and how they might be regulated by outside agents. Collaborations on cellular aspects on these receptors with Drs. Andrei Gudkov, Cynthia Leifer and Jenny Ting and on specialized biophysical approaches with Drs. Andrew Ward and David Baker will complement our biophysical and functional studies as well as stimulate development of agonists and antagonists as therapeutics in compromised immunity or in hyper- inflammatory diseases.

Public Health Relevance

Structural, biophysical and cellular studies of innate immunity will focus on Toll-like receptors (TLRs) and NOD-like receptors (NLRs). Structures of leucine-rich repeats (LRRs) in TLRs and NLRs by themselves or complexed with their signaling partners will advance our understanding of receptor activation and regulatory mechanisms in innate immunity. Such findings will aid in the development of agonists and antagonists as adjuvants, or as therapeutics in compromised immunity or in hyper-inflammatory diseases, respectively.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
5R01AI042266-17
Application #
8606135
Study Section
Macromolecular Structure and Function C Study Section (MSFC)
Program Officer
Leitner, Wolfgang W
Project Start
1998-01-01
Project End
2018-01-31
Budget Start
2014-02-01
Budget End
2015-01-31
Support Year
17
Fiscal Year
2014
Total Cost
$544,210
Indirect Cost
$257,028
Name
Scripps Research Institute
Department
Type
DUNS #
781613492
City
La Jolla
State
CA
Country
United States
Zip Code
92037
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