title: Multiple molecular regions confer intracellular localization of bovine Toll-like receptor 8.
abstract: Toll-like receptors (TLRs), a family of highly conserved germline-encoded pattern-recognition receptors (PRRs), are essential for the host immune response. The cellular localization of TLR proteins determines the access to certain sources of ligands and thus the triggering of downstream cellular signaling. The TLR7/8/9 subfamily proteins are localized intracellularly but the molecular elements determining the cellular localization of these proteins are not fully understood. Here we demonstrated that the bovine TLR8 (bTLR8) protein is localized in the ER cellular compartment of transfected cells before and after cell activation. Using chimeric constructs, we showed that the bTLR8 transmembrane (TM) and cytoplasmic (CP) regions could direct the bovine herpesvirus 1 (BHV-1) glycoprotein D (gD) extracellular domain (ECD) to an intracellular localization. Furthermore, the bTLR8 TM, the linker region between the TM and TIR domains, and the TIR-tail region all partially contributed to the intracellular localization. However, truncation of the bTLR8 with the TM and CP regions removed did not alter its intracellular localization, suggesting that ectodomain (ECD) itself contains intracellular information. Indeed, the bTLR8 ECD also targeted the gD ECD to the intracellular localization. Our results suggest that multiple regions, including ECD, TM, linker and TIR-tail regions of bTLR8, are involved in determining the localization of cellular ER compartment.
authors.affiliation: Vaccine and Infectious Disease Organization (VIDO), University of Saskatchewan, Saskatoon, Saskatchewan, Canada S7N 5E3.
authors.forename: Jianzhong
authors.initials: J
authors.lastname: Zhu
authors.forename: Sylvia
authors.initials: S
authors.lastname: van Drunen Littel-van den Hurk
authors.forename: Robert
authors.initials: R
authors.lastname: Brownlie
authors.forename: Lorne A
authors.initials: LA
authors.lastname: Babiuk
authors.forename: Andrew
authors.initials: A
authors.lastname: Potter
authors.forename: George K
authors.initials: GK
authors.lastname: Mutwiri
mesh_terms: D000818:Animals; D019556:COS Cells; D002417:Cattle / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D002522:Chlorocebus aethiops; D004721:Endoplasmic Reticulum / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D010449:Peptide Mapping / Q000379:methods; D017434:Protein Structure, Tertiary / Q000502:physiology; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D015398:Signal Transduction / Q000502:physiology*; D051200:Toll-Like Receptor 8 / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D014764:Viral Proteins / Q000235:genetics / Q000276:immunology / Q000378:metabolism
chemical_list: D011993:Recombinant Fusion Proteins; D051200:Toll-Like Receptor 8; D014764:Viral Proteins; C046998:bovine herpesvirus type-1 glycoproteins