title: The polymorphic HCMV glycoprotein UL20 is targeted for lysosomal degradation by multiple cytoplasmic dileucine motifs.
abstract: Human cytomegalovirus (HCMV) is a widespread and persistent beta-herpesvirus. The large DNA genome of HCMV encodes many proteins that are non-essential for viral replication including numerous proteins subverting host immunosurveillance. One of them is the barely characterized UL20, which is encoded adjacent to the well-defined immunoevasins UL16 and UL18. UL20 is a type I transmembrane glycoprotein with an immunoglobulin-like ectodomain that is highly polymorphic among HCMV strains. Here, we show that the homodimeric UL20, by virtue of its cytoplasmic domain, does not reach the cell surface but is targeted to endosomes and lysosomes. Accordingly, UL20 exhibits a short half-life because of rapid lysosomal degradation. Trafficking of UL20 to lysosomes is determined by several, independently functioning dileucine-based sorting motifs in the cytoplasmic domain of UL20 and involves the adaptor protein (AP) complex AP-1. Combined substitution of three dileucine motifs allowed strong cell surface expression of UL20 comparable to UL20 mutants lacking the cytoplasmic tail. Finally, we show that the intracellularly located UL20 also is subject to lysosomal degradation in the context of viral infection. Altogether, from these data, we hypothesize that UL20 is destined to efficiently sequester yet-to-be defined cellular proteins for degradation in lysosomes.
authors.affiliation: Institute for Molecular Medicine, Goethe-University Frankfurt am Main, 60590 Frankfurt am Main, Germany.
authors.forename: Ivan
authors.initials: I
authors.lastname: Jelcic
authors.forename: Johanna
authors.initials: J
authors.lastname: Reichel
authors.forename: Christoph
authors.initials: C
authors.lastname: Schlude
authors.forename: Eva
authors.initials: E
authors.lastname: Treutler
authors.forename: Christian
authors.initials: C
authors.lastname: Sinzger
authors.forename: Alexander
authors.initials: A
authors.lastname: Steinle
mesh_terms: D033961:Adaptor Protein Complex 1 / Q000378:metabolism; D020816:Amino Acid Motifs; D002460:Cell Line; D003587:Cytomegalovirus / Q000235:genetics / Q000254:growth & development / Q000378:metabolism*; D003593:Cytoplasm / Q000378:metabolism*; D004591:Electrophoresis, Polyacrylamide Gel; D005347:Fibroblasts / Q000821:virology; D005434:Flow Cytometry; D006801:Humans; D007930:Leucine / Q000378:metabolism*; D008247:Lysosomes / Q000378:metabolism*; D018942:Macrolides / Q000494:pharmacology; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D021381:Protein Transport; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D014162:Transfection; D014764:Viral Proteins / Q000235:genetics / Q000378:metabolism*
chemical_list: D033961:Adaptor Protein Complex 1; D018942:Macrolides; D011993:Recombinant Fusion Proteins; D014764:Viral Proteins; C040929:bafilomycin A1; D007930:Leucine