title: A pH- and ionic strength-dependent conformational change in the neck region regulates DNGR-1 function in dendritic cells.
abstract: DNGR-1 is receptor expressed by certain dendritic cell (DC) subsets and by DC precursors in mouse. It possesses a C-type lectin-like domain (CTLD) followed by a poorly characterized neck region coupled to a transmembrane region and short intracellular tail. The CTLD of DNGR-1 binds F-actin exposed by dead cell corpses and causes the receptor to signal and potentiate cross-presentation of dead cell-associated antigens by DCs. Here, we describe a conformational change that occurs in the neck region of DNGR-1 in a pH- and ionic strength-dependent manner and that controls cross-presentation of dead cell-associated antigens. We identify residues in the neck region that, when mutated, lock DNGR-1 in one of the two conformational states to potentiate cross-presentation. In contrast, we show that chimeric proteins in which the neck region of DNGR-1 is replaced by that of unrelated C-type lectin receptors fail to promote cross-presentation. Our results suggest that the neck region of DNGR-1 is an integral receptor component that senses receptor progression through the endocytic pathway and has evolved to maximize extraction of antigens from cell corpses, coupling DNGR-1 function to its cellular localization.
authors.affiliation: Immunobiology Laboratory, The Francis Crick Institute, London, UK.
authors.forename: Pavel
authors.initials: P
authors.lastname: Hanč
authors.affiliation: Immunobiology Laboratory, The Francis Crick Institute, London, UK.
authors.forename: Oliver
authors.initials: O
authors.lastname: Schulz
authors.affiliation: Immunobiology Laboratory, The Francis Crick Institute, London, UK.
authors.forename: Hanna
authors.initials: H
authors.lastname: Fischbach
authors.affiliation: Structural Biology Science Technology Platform, The Francis Crick Institute, London, UK.
authors.forename: Stephen R
authors.initials: SR
authors.lastname: Martin
authors.affiliation: Structural Biology Science Technology Platform, The Francis Crick Institute, London, UK.
authors.forename: Svend
authors.initials: S
authors.lastname: Kjær
authors.affiliation: Immunobiology Laboratory, The Francis Crick Institute, London, UK caetano@crick.ac.uk.
authors.forename: Caetano
authors.identifier: 0000-0001-7392-2119
authors.initials: C
authors.lastname: Reis e Sousa
mesh_terms: D000494:Allosteric Regulation; D000818:Animals; D004252:DNA Mutational Analysis; D003713:Dendritic Cells / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration; D037181:Lectins, C-Type / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D051379:Mice; D009994:Osmolar Concentration; D011487:Protein Conformation / Q000187:drug effects*; D011971:Receptors, Immunologic / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*
chemical_list: C529914:Clec9a protein, mouse; D037181:Lectins, C-Type; D011971:Receptors, Immunologic
keywords: C‐type lectin receptors; cross‐presentation; necrosis; protein structure