title: Nucleic acid recognition by Toll-like receptors is coupled to stepwise processing by cathepsins and asparagine endopeptidase.
abstract: Toll-like receptor (TLR) 9 requires proteolytic processing in the endolysosome to initiate signaling in response to DNA. However, recent studies conflict as to which proteases are required for receptor cleavage. We show that TLR9 proteolysis is a multistep process. The first step removes the majority of the ectodomain and can be performed by asparagine endopeptidase (AEP) or cathepsin family members. This initial cleavage event is followed by a trimming event that is solely cathepsin mediated and required for optimal receptor signaling. This dual requirement for AEP and cathepsins is observed in all cell types that we have analyzed, including mouse macrophages and dendritic cells. In addition, we show that TLR7 and TLR3 are processed in an analogous manner. These results define the core proteolytic steps required for TLR9 function and suggest that receptor proteolysis may represent a general regulatory strategy for all TLRs involved in nucleic acid recognition.
authors.affiliation: Division of Immunology and Pathogenesis, Department of Molecular and Cell Biology, University of California, Berkeley, Berkeley, CA 94720, USA.
authors.forename: Sarah E
authors.initials: SE
authors.lastname: Ewald
authors.forename: Alex
authors.initials: A
authors.lastname: Engel
authors.forename: Jiyoun
authors.initials: J
authors.lastname: Lee
authors.forename: Miqi
authors.initials: M
authors.lastname: Wang
authors.forename: Matthew
authors.initials: M
authors.lastname: Bogyo
authors.forename: Gregory M
authors.initials: GM
authors.lastname: Barton
mesh_terms: D000818:Animals; D002403:Cathepsins / Q000502:physiology*; D002460:Cell Line; D003546:Cysteine Endopeptidases / Q000502:physiology*; D003713:Dendritic Cells / Q000378:metabolism; D008562:Membrane Glycoproteins / Q000378:metabolism; D051379:Mice; D008810:Mice, Inbred C57BL; D009696:Nucleic Acids / Q000378:metabolism*; D051196:Toll-Like Receptor 3 / Q000378:metabolism; D051199:Toll-Like Receptor 7 / Q000378:metabolism; D051217:Toll-Like Receptor 9 / Q000378:metabolism*; D000099240:Asparaginyl Endopeptidase
chemical_list: D002403:Cathepsins; D003546:Cysteine Endopeptidases; D008562:Membrane Glycoproteins; D009696:Nucleic Acids; D051196:Toll-Like Receptor 3; D051199:Toll-Like Receptor 7; D051217:Toll-Like Receptor 9; D000099240:Asparaginyl Endopeptidase; C519703:TLR3 protein, mouse; C477038:Tlr7 protein, mouse; C495370:Tlr9 protein, mouse