Ligand-specific, transient interaction between integrins and calreticulin during cell adhesion to extracellular matrix proteins is dependent upon phosphorylation dephosphorylation events

被引:56
作者
Coppolino, MG
Dedhar, S
机构
[1] Hosp Sick Children, Dept Cell Biol, Toronto, ON M5G 1X8, Canada
[2] British Columbia Canc Agcy, Jack Bell Res Ctr, Vancouver, BC V6H 3Z6, Canada
关键词
cell attachment; early adhesion complex; MAP kinase; protein phosphatase; signal transduction;
D O I
10.1042/0264-6021:3400041
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
As transmembrane heterodimers, integrins bind to both extracellular ligands and intracellular proteins. We are currently investigating the interaction between integrins and the intracellular protein calreticulin. A prostatic carcinoma cell line (PC-3) was used to demonstrate that calreticulin can be found in the alpha(3) immunoprecipitates of cells plated on collagen type IV, but not when plated on vitronectin. Conversely, alpha(v) immunoprecipitates contained calreticulin only when cells were plated on vitronectin, i.e. not when plated on collagen IV. The interactions between these integrins and calreticulin were independent of actin cytoskeleton assembly and were transient, being maximal approx. 10-30 min after the cells came into contact with the substrates prior to complete cell spreading and formation of firm adhesive contacts. We demonstrate that okadaic acid, an inhibitor of intracellular serine/threonine protein phosphatases, inhibited the alpha(3)beta(1)-mediated adhesion of PC-3 cells to collagen EV and the alpha(2)beta(1)-mediated attachment of Jurkat cells to collagen I. This inhibition by okadaic acid was accompanied by inhibition of the ligand-specific interaction of calreticulin with the respective integrins in the two cell types. Additionally, we found that pharmacological inhibition of mitogen-activated protein kinase kinase (MEK) resulted in prolongation of the calreticulin-integrin interaction, and enhancement of PC-3 cell attachment to collagen IV. We conclude that calreticulin interacts transiently with integrins during cell attachment and spreading. This interaction depends on receptor occupation, is ligand-specific, and can be modulated by protein phosphatase and MEK activity.
引用
收藏
页码:41 / 50
页数:10
相关论文
共 49 条
  • [1] PD-098059 IS A SPECIFIC INHIBITOR OF THE ACTIVATION OF MITOGEN-ACTIVATED PROTEIN-KINASE KINASE IN-VITRO AND IN-VIVO
    ALESSI, DR
    CUENDA, A
    COHEN, P
    DUDLEY, DT
    SALTIEL, AR
    [J]. JOURNAL OF BIOLOGICAL CHEMISTRY, 1995, 270 (46) : 27489 - 27494
  • [2] BARTFELD NS, 1993, J BIOL CHEM, V268, P17270
  • [3] Characterization of novel complexes on the cell surface between integrins and proteins with 4 transmembrane domains (TM4 proteins)
    Berditchevski, F
    Zutter, MM
    Hemler, ME
    [J]. MOLECULAR BIOLOGY OF THE CELL, 1996, 7 (02) : 193 - 207
  • [4] SPECIFIC ASSOCIATION OF CD63 WITH THE VLA-3 AND VLA-6 INTEGRINS
    BERDITCHEVSKI, F
    BAZZONI, G
    HEMLER, ME
    [J]. JOURNAL OF BIOLOGICAL CHEMISTRY, 1995, 270 (30) : 17784 - 17790
  • [5] INTEGRIN-ASSOCIATED PROTEIN - A 50-KD PLASMA-MEMBRANE ANTIGEN PHYSICALLY AND FUNCTIONALLY ASSOCIATED WITH INTEGRINS
    BROWN, E
    HOOPER, L
    HO, T
    GRESHAM, H
    [J]. JOURNAL OF CELL BIOLOGY, 1990, 111 (06) : 2785 - 2794
  • [6] MODULATION OF GENE-EXPRESSION BY CALRETICULIN BINDING TO THE GLUCOCORTICOID RECEPTOR
    BURNS, K
    DUGGAN, B
    ATKINSON, EA
    FAMULSKI, KS
    NEMER, M
    BLEACKLEY, RC
    MICHALAK, M
    [J]. NATURE, 1994, 367 (6462) : 476 - 480
  • [7] ICAP-1, a novel beta(1) integrin cytoplasmic domain-associated protein, binds to a conserved and functionally important NPXY sequence motif of beta(1) integrin
    Chang, DD
    Wong, C
    Smith, H
    Liu, J
    [J]. JOURNAL OF CELL BIOLOGY, 1997, 138 (05) : 1149 - 1157
  • [8] INDUCIBLE INTERACTION OF INTEGRIN ALPHA(2)BETA(1) WITH CALRETICULIN - DEPENDENCE ON THE ACTIVATION STATE OF THE INTEGRIN
    COPPOLINO, M
    LEUNGHAGESTEIJN, C
    DEDHAR, S
    WILKINS, J
    [J]. JOURNAL OF BIOLOGICAL CHEMISTRY, 1995, 270 (39) : 23132 - 23138
  • [9] Calreticulin is essential for integrin-mediated calcium signalling and cell adhesion
    Coppolino, MG
    Woodside, MJ
    Demaurex, N
    Grinstein, S
    StArnaud, R
    Dedhar, S
    [J]. NATURE, 1997, 386 (6627) : 843 - 847
  • [10] Calreticulin
    Coppolino, MG
    Dedhar, S
    [J]. INTERNATIONAL JOURNAL OF BIOCHEMISTRY & CELL BIOLOGY, 1998, 30 (05) : 553 - 558