B Cell Receptor-induced Phosphorylation of Pyk2 and Focal Adhesion Kinase Involves Integrins and the Rap GTPases and Is Required for B Cell Spreading

被引:38
作者
Tse, Kathy W. K. [1 ,2 ,4 ]
Dang-Lawson, May [1 ,2 ,4 ]
Lee, Rosaline L. [1 ,2 ,4 ]
Vong, Doris [1 ,2 ,4 ]
Bulic, Anica [1 ,2 ,4 ]
Buckbinder, Leonard [3 ]
Gold, Michael R. [1 ,2 ,4 ]
机构
[1] Univ British Columbia, Dept Microbiol & Immunol, Vancouver, BC V6T 1Z3, Canada
[2] Univ British Columbia, Inst Life Sci, I3 Res Grp, Vancouver, BC V6T 1Z3, Canada
[3] Pfizer Global Res & Dev, Groton, CT 06340 USA
[4] Univ British Columbia, Inst Life Sci, CELL Res Grp, Vancouver, BC V6T 1Z3, Canada
基金
加拿大健康研究院; 加拿大自然科学与工程研究理事会;
关键词
MEDIATES RAP1-INDUCED ADHESION; RICH TYROSINE KINASE-2; CYTOSKELETAL REARRANGEMENT; ACTIN POLYMERIZATION; INDUCED ACTIVATION; SYNAPSE FORMATION; SIGNALING EVENTS; MIGRATION; PROTEIN; LYMPHOCYTES;
D O I
10.1074/jbc.M109.013169
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Signaling by the B cell receptor (BCR) promotes integrin-mediated adhesion and cytoskeletal reorganization. This results in B cell spreading, which enhances the ability of B cells to bind antigens and become activated. Proline-rich tyrosine kinase (Pyk2) and focal adhesion kinase (FAK) are related cytoplasmic tyrosine kinases that regulate cell adhesion, cell morphology, and cell migration. In this report we show that BCR signaling and integrin signaling collaborate to induce the phosphorylation of Pyk2 and FAK on key tyrosine residues, a modification that increases the kinase activity of Pyk2 and FAK. Activation of the Rap GTPases is critical for BCR-induced integrin activation as well as for BCR- and integrin-induced reorganization of the actin cytoskeleton. We now show that Rap activation is essential for BCR- induced phosphorylation of Pyk2 and for integrin-induced phosphorylation of Pyk2 and FAK. Moreover Rap-dependent phosphorylation of Pyk2 and FAK required an intact actin cytoskeleton as well as actin dynamics, suggesting that Rap regulates Pyk2 and FAK via its effects on the actin cytoskeleton. Importantly B cell spreading induced by BCR/integrin co-stimulation or by integrin engagement was inhibited by short hairpin RNA-mediated knockdown of either Pyk2 or FAK expression and by treatment with PF-431396, a chemical inhibitor that blocks the kinase activities of both Pyk2 and FAK. Thus Pyk2 and FAK are downstream targets of the Rap GTPases that play a key role in regulating B cell morphology.
引用
收藏
页码:22865 / 22877
页数:13
相关论文
共 69 条
[1]   Distinct types of diffuse large B-cell lymphoma identified by gene expression profiling [J].
Alizadeh, AA ;
Eisen, MB ;
Davis, RE ;
Ma, C ;
Lossos, IS ;
Rosenwald, A ;
Boldrick, JG ;
Sabet, H ;
Tran, T ;
Yu, X ;
Powell, JI ;
Yang, LM ;
Marti, GE ;
Moore, T ;
Hudson, J ;
Lu, LS ;
Lewis, DB ;
Tibshirani, R ;
Sherlock, G ;
Chan, WC ;
Greiner, TC ;
Weisenburger, DD ;
Armitage, JO ;
Warnke, R ;
Levy, R ;
Wilson, W ;
Grever, MR ;
Byrd, JC ;
Botstein, D ;
Brown, PO ;
Staudt, LM .
NATURE, 2000, 403 (6769) :503-511
[2]   Imaging of germinal center selection events during affinity maturation [J].
Allen, Christopher D. C. ;
Okada, Takaharu ;
Tang, H. Lucy ;
Cyster, Jason G. .
SCIENCE, 2007, 315 (5811) :528-531
[3]   α6-integrin is expressed on germinal centre B cells and modifies growth of a B-cell line [J].
Ambrose, HE ;
Wagner, SD .
IMMUNOLOGY, 2004, 111 (04) :400-406
[4]  
Andreev J, 1999, MOL CELL BIOL, V19, P2338
[5]  
Astier A, 1997, J BIOL CHEM, V272, P228
[6]   RAFTK/Pyk2-mediated cellular signalling [J].
Avraham, H ;
Park, SY ;
Schinkmann, K ;
Avraham, S .
CELLULAR SIGNALLING, 2000, 12 (03) :123-133
[7]   Distinct roles for LFA-1 and CD28 during activation of naive T cells: Adhesion versus costimulation [J].
Bachmann, MF ;
McKallFaienza, K ;
Schmits, R ;
Bouchard, D ;
Beach, J ;
Speiser, DE ;
Mak, TW ;
Ohashi, PS .
IMMUNITY, 1997, 7 (04) :549-557
[8]   Stromal cell networks regulate lymphocyte entry, migration, and territoriality in lymph nodes [J].
Bajenoff, Marc ;
Egen, Jackson G. ;
Koo, Lily Y. ;
Laugier, Jean Pierre ;
Brau, Frederic ;
Glaichenhaus, Nicolas ;
Germain, Ronald N. .
IMMUNITY, 2006, 25 (06) :989-1001
[9]   Cell surface recycling of internalized antigen permits dendritic cell priming of B cells [J].
Bergtold, A ;
Desai, DD ;
Gavhane, A ;
Clyne, R .
IMMUNITY, 2005, 23 (05) :503-514
[10]   Linking Rap to cell adhesion [J].
Bos, JL .
CURRENT OPINION IN CELL BIOLOGY, 2005, 17 (02) :123-128