Enrichment of Lck in lipid rafts regulates colocalized Fyn activation and the initiation of proximal signals through TCRαβ

被引:35
作者
Filipp, D
Leung, BL
Zhang, J
Veillette, A
Julius, M
机构
[1] Univ Toronto, Sunnybrook & Womens Coll Hlth Sci Ctr, Toronto, ON, Canada
[2] Univ Toronto, Dept Immunol, Toronto, ON, Canada
[3] McGill Univ, Mol Oncol Lab, Inst Rech Clin Montreal, Montreal, PQ, Canada
[4] McGill Univ, Dept Microbiol & Immunol, Montreal, PQ, Canada
关键词
D O I
10.4049/jimmunol.172.7.4266
中图分类号
R392 [医学免疫学]; Q939.91 [免疫学];
学科分类号
100102 [免疫学];
摘要
Recent results provide insight into the temporal and spatial relationship governing lck-dependent fyn activation and demonstrate TCR/CD4-induced activation and translocation of lck into lipid rafts and the ensuing activation of colocalized fyn. The prediction follows that directly targeting lck to lipid rafts will bypass the requirement for juxtaposing TCR and CD4-lck, and rescue cellular activation mediated by Ab specific for the constant region of TCRbeta chain. The present study uses a family of murine IL-2-dependent CD4(+) T cell clonal variants in which anti-TCRCbeta signaling is impaired in an lck-dependent fashion. Importantly, these variants respond to Ag- and mAb-mediated TCR-CD4 coaggregation, both of which enable the coordinated interaction of CD4-associated lck with the TCR/CD3 complex. We have previously demonstrated that anti-TCRCbeta responsiveness in this system correlates with the presence of kinase-active, membrane-associated lck and preformed hypophosphorylated TCRzeta:zeta-associated protein of 70 kDa complexes, a phenotype recapitulated in primary resting CD4(+) T cells. We show in this study that forced expression of wild-type lck achieved the same basal composition of the TCR/CD3 complex and yet did not rescue anti-TCRCbeta signaling. In contrast, forced expression of C20S/C23S-mutated lck (double-cysteine lck), unable to bind CD4, rescues anti-TCRCbeta proximal signaling and cellular growth. Double-cysteine lck targets lipid rafts, colocalizes with >98% of cellular fyn, and results in a 7-fold increase in basal fyn kinase activity. Coaggregation of CD4 and TCR achieves the same outcome. These results underscore the critical role of lipid rafts in spatially coordinating the interaction between lck and fyn that predicates proximal TCR/CD3 signaling.
引用
收藏
页码:4266 / 4274
页数:9
相关论文
共 71 条
[1]
ENHANCEMENT OF T-CELL RESPONSIVENESS BY THE LYMPHOCYTE-SPECIFIC TYROSINE PROTEIN-KINASE P56LCK [J].
ABRAHAM, N ;
MICELI, MC ;
PARNES, JR ;
VEILLETTE, A .
NATURE, 1991, 350 (6313) :62-66
[2]
Distinct patterns of membrane microdomain partitioning in Th1 and Th2 cells [J].
Balamuth, F ;
Leitenberg, D ;
Unternaehrer, J ;
Mellman, I ;
Bottomly, K .
IMMUNITY, 2001, 15 (05) :729-738
[3]
INTERACTIONS BETWEEN THE TYROSINE KINASES P56LCK, P59FYN AND P50CSK IN CD4 SIGNALING IN T-CELLS [J].
BALDARI, CT ;
DISOMMA, MM ;
MILIA, E ;
BERGMAN, M ;
TELFORD, JL .
EUROPEAN JOURNAL OF IMMUNOLOGY, 1995, 25 (04) :919-925
[4]
Phosphoprotein associated with glycosphingolipid-enriched microdomains (PAG), a novel ubiquitously expressed transmembrane adaptor protein, binds the protein tyrosine kinase Csk and is involved in regulation of T cell activation [J].
Brdicka, T ;
Pavilstová, D ;
Leo, A ;
Bruyns, E ;
Korínek, V ;
Angelisová, P ;
Scherer, J ;
Shevchenko, A ;
Shevchenko, A ;
Hilgert, I ;
Cerny, J ;
Drbal, K ;
Kuramitsu, Y ;
Kornacker, B ;
Horejsí, V ;
Schraven, B .
JOURNAL OF EXPERIMENTAL MEDICINE, 2000, 191 (09) :1591-1604
[5]
The immunological synapse [J].
Bromley, SK ;
Burack, WR ;
Johnson, KG ;
Somersalo, K ;
Sims, TN ;
Sumen, C ;
Davis, MM ;
Shaw, AS ;
Allen, PM ;
Dustin, ML .
ANNUAL REVIEW OF IMMUNOLOGY, 2001, 19 :375-396
[6]
ACTIVATION OF ZAP-70 KINASE-ACTIVITY BY PHOSPHORYLATION OF TYROSINE-493 IS REQUIRED FOR LYMPHOCYTE ANTIGEN RECEPTOR FUNCTION [J].
CHAN, AC ;
DALTON, M ;
JOHNSON, R ;
KONG, GH ;
WANG, T ;
THOMA, R ;
KUROSAKI, T .
EMBO JOURNAL, 1995, 14 (11) :2499-2508
[7]
ZAP-70 - A 70 KD PROTEIN-TYROSINE KINASE THAT ASSOCIATES WITH THE TCR ZETA-CHAIN [J].
CHAN, AC ;
IWASHIMA, M ;
TURCK, CW ;
WEISS, A .
CELL, 1992, 71 (04) :649-662
[8]
Cloning of a novel T-cell protein FYB that binds FYN and SH2-domain-containing leukocyte protein 76 and modulates interleukin 2 production [J].
DaSilva, AJ ;
Li, ZW ;
DeVera, C ;
Canto, E ;
Findell, P ;
Rudd, CE .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1997, 94 (14) :7493-7498
[9]
Phosphorylation-dependent regulation of T-cell activation by PAG/Cbp, a lipid raft-associated transmembrane adaptor [J].
Davidson, D ;
Bakinowski, M ;
Thomas, ML ;
Horejsi, V ;
Veillette, A .
MOLECULAR AND CELLULAR BIOLOGY, 2003, 23 (06) :2017-2028
[10]
DIFFERENTIAL REGULATION OF T-CELL ANTIGEN RESPONSIVENESS BY ISOFORMS OF THE SRC-RELATED TYROSINE PROTEIN-KINASE P59FYN [J].
DAVIDSON, D ;
CHOW, LML ;
FOURNEL, M ;
VEILLETTE, A .
JOURNAL OF EXPERIMENTAL MEDICINE, 1992, 175 (06) :1483-1492