title: The variable hinge region of novel PKCs determines localization to distinct regions of the immunological synapse.
abstract: The immunological synapse (IS) formed between a T cell and its cognate antigen-presenting cell (APC) enables the directional secretion of cytolytic and inflammatory molecules. Synaptic architecture is established in part by a two-step cascade of novel protein kinase C (nPKC) isozymes. PKCε and PKCη arrive at the IS first, and occupy the entire synaptic membrane. Then, PKCθ accumulates in a smaller zone at the center of the contact. We investigated the molecular basis for this differential recruitment behavior using chimeric nPKC constructs and total internal reflection fluorescence microscopy. Our studies revealed that the V3 linker just N-terminal to the kinase domain plays a crucial role in specifying nPKC localization. Substitution of this linker switched the scope and the kinetics of PKCθ accumulation to that of PKCε and PKCη, and vice versa. Although the V3 was necessary for synaptic compartmentalization, it was not sufficient, as the tandem C1 domains were also required to mediate membrane association. Together, these results suggest a model whereby the V3 linker controls nPKC sub-compartmentalization after initial C1 domain-mediated accumulation at the IS.
authors.affiliation: Immunology Program, Memorial Sloan-Kettering Cancer Center, New York, New York, United States of America.
authors.forename: Roshni
authors.initials: R
authors.lastname: Basu
authors.affiliation: Immunology Program, Memorial Sloan-Kettering Cancer Center, New York, New York, United States of America.
authors.forename: Yuedan
authors.initials: Y
authors.lastname: Chen
authors.affiliation: Immunology Program, Memorial Sloan-Kettering Cancer Center, New York, New York, United States of America.
authors.forename: Emily J
authors.initials: EJ
authors.lastname: Quann
authors.affiliation: Immunology Program, Memorial Sloan-Kettering Cancer Center, New York, New York, United States of America.
authors.forename: Morgan
authors.initials: M
authors.lastname: Huse
mesh_terms: D000818:Animals; D054992:Immunological Synapses / Q000378:metabolism*; D007700:Kinetics; D051379:Mice; D011493:Protein Kinase C / Q000737:chemistry* / Q000378:metabolism*; D017434:Protein Structure, Tertiary; D021381:Protein Transport; D013329:Structure-Activity Relationship
chemical_list: D011493:Protein Kinase C
references.citation: Sci Signal. 2011 Dec 06;4(202):ra84
references.pmid: 22155788
references.citation: Nat Immunol. 2009 Jun;10(6):627-35
references.pmid: 19430478
references.citation: Nat Immunol. 2011 Oct 02;12(11):1105-12
references.pmid: 21964608
references.citation: Immunity. 2007 Jul;27(1):76-88
references.pmid: 17629516
references.citation: Cold Spring Harb Perspect Biol. 2010 Oct;2(10):a002311
references.pmid: 20843980
references.citation: Nat Immunol. 2012 Jul 01;13(8):787-95
references.pmid: 22751140
references.citation: Mol Biol Cell. 2004 Jun;15(6):2932-42
references.pmid: 15064353
references.citation: Immunity. 2006 May;24(5):535-46
references.pmid: 16713972
references.citation: Front Immunol. 2012 Jul 31;3:235
references.pmid: 23060874
references.citation: J Exp Med. 2013 Nov 18;210(12):2721-37
references.pmid: 24190432
references.citation: Cell. 2007 May 18;129(4):773-85
references.pmid: 17512410
references.citation: J Biol Chem. 2005 May 20;280(20):19784-93
references.pmid: 15769752
references.citation: Science. 2010 Apr 16;328(5976):372-6
references.pmid: 20339032
references.citation: Am J Physiol Endocrinol Metab. 2010 Mar;298(3):E395-402
references.pmid: 19934406
references.citation: Mol Biol Cell. 2011 Sep;22(18):3491-7
references.pmid: 21795397
references.citation: J Biol Chem. 2007 Jul 20;282(29):21467-76
references.pmid: 17548359
references.citation: Immunity. 2008 Oct 17;29(4):589-601
references.pmid: 18848472
references.citation: Nat Immunol. 2011 May 22;12(7):647-54
references.pmid: 21602810
references.citation: J Immunol. 2012 Nov 15;189(10):4859-69
references.pmid: 23077238
references.citation: Science. 2005 Nov 18;310(5751):1191-3
references.pmid: 16293763
references.citation: Curr Opin Cell Biol. 2009 Apr;21(2):262-7
references.pmid: 19195860