title: Varied Role of Ubiquitylation in Generating MHC Class I Peptide Ligands.
abstract: CD8+T cell immunosurveillance is based on recognizing oligopeptides presented by MHC class I molecules. Despite decades of study, the importance of protein ubiquitylation to peptide generation remains uncertain. In this study, we examined the ability of MLN7243, a recently described ubiquitin-activating enzyme E1 inhibitor, to block overall cytosolic peptide generation and generation of specific peptides from vaccinia- and influenza A virus-encoded proteins. We show that MLN7243 rapidly inhibits ubiquitylation in a variety of cell lines and can profoundly reduce the generation of cytosolic peptides. Kinetic analysis of specific peptide generation reveals that ubiquitylation of defective ribosomal products is rate limiting in generating class I peptide complexes. More generally, our findings demonstrate that the requirement for ubiquitylation in MHC class I-restricted Ag processing varies with class I allomorph, cell type, source protein, and peptide context. Thus, ubiquitin-dependent and -independent pathways robustly contribute to MHC class I-based immunosurveillance.
authors.affiliation: Laboratory of Viral Diseases, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, MD 20892.
authors.forename: Jiajie
authors.initials: J
authors.lastname: Wei
authors.affiliation: T Cell Laboratory, School of Molecular Science, La Trobe University, Bundoora, Victoria 3086, Australia.
authors.forename: Damien
authors.identifier: 0000-0003-1163-9902
authors.initials: D
authors.lastname: Zanker
authors.affiliation: T Cell Laboratory, School of Molecular Science, La Trobe University, Bundoora, Victoria 3086, Australia.
authors.forename: Anthony R
authors.identifier: 0000-0002-6925-3829
authors.initials: AR
authors.lastname: Di Carluccio
authors.affiliation: Biological Imaging Section, Research Technologies Branch, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, MD 20892.
authors.forename: Margery G
authors.identifier: 0000-0001-7777-5574
authors.initials: MG
authors.lastname: Smelkinson
authors.affiliation: Microscopy and Imaging Core Facility, Center for Biologics Evaluation and Research, Food and Drug Administration, Silver Spring, MD 20993; and.
authors.forename: Kazuyo
authors.initials: K
authors.lastname: Takeda
authors.affiliation: Laboratory of Viral Diseases, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, MD 20892.
authors.forename: Mina O
authors.initials: MO
authors.lastname: Seedhom
authors.affiliation: Laboratory of Viral Diseases, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, MD 20892.
authors.forename: Devin
authors.initials: D
authors.lastname: Dersh
authors.affiliation: Laboratory of Viral Diseases, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, MD 20892.
authors.forename: James S
authors.initials: JS
authors.lastname: Gibbs
authors.affiliation: Laboratory of Viral Diseases, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, MD 20892.
authors.forename: Ning
authors.identifier: 0000-0001-5121-7208
authors.initials: N
authors.lastname: Yang
authors.affiliation: Chemical Genomics Center, National Center for Advancing Translational Sciences, National Institutes of Health, Rockville, MD 20892.
authors.forename: Ajit
authors.initials: A
authors.lastname: Jadhav
authors.affiliation: T Cell Laboratory, School of Molecular Science, La Trobe University, Bundoora, Victoria 3086, Australia.
authors.forename: Weisan
authors.identifier: 0000-0002-5221-9771
authors.initials: W
authors.lastname: Chen
authors.affiliation: Laboratory of Viral Diseases, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, MD 20892; jyewdell@niaid.nih.gov.
authors.forename: Jonathan W
authors.identifier: 0000-0003-2123-6189
authors.initials: JW
authors.lastname: Yewdell
mesh_terms: D000818:Animals; D017951:Antigen Presentation; D002460:Cell Line; D003600:Cytosol / Q000737:chemistry / Q000276:immunology; D004791:Enzyme Inhibitors / Q000494:pharmacology; D015395:Histocompatibility Antigens Class I / Q000276:immunology* / Q000378:metabolism; D006801:Humans; D009980:Influenza A virus / Q000737:chemistry / Q000276:immunology; D007700:Kinetics; D008024:Ligands; D051379:Mice; D015166:Monitoring, Immunologic; D009705:Nucleosides / Q000494:pharmacology*; D010455:Peptides / Q000276:immunology* / Q000378:metabolism; D011720:Pyrazoles; D011743:Pyrimidines; D013440:Sulfides; D013449:Sulfonamides / Q000494:pharmacology*; D013601:T-Lymphocytes / Q000276:immunology*; D054875:Ubiquitination; D014616:Vaccinia virus / Q000737:chemistry / Q000276:immunology
chemical_list: D004791:Enzyme Inhibitors; D015395:Histocompatibility Antigens Class I; D008024:Ligands; D009705:Nucleosides; D010455:Peptides; D011720:Pyrazoles; D011743:Pyrimidines; D013440:Sulfides; D013449:Sulfonamides; C000622638:TAK-243