Real time fluorescence imaging of PLCγ translocation and its interaction with the epidermal growth factor receptor

被引:67
作者
Matsuda, M
Paterson, HF
Rodriguez, R
Fensome, AC
Ellis, MV
Swann, K
Katan, M
机构
[1] Inst Canc Res, CRC, Ctr Cell & Mol Biol, Chester Beatty Labs, London SW3 6JB, England
[2] UCL, Dept Anat & Dev Biol, London WC1E 6BT, England
关键词
PLC; EGF receptor; translocation; real time imaging; SH2 and PH domains;
D O I
10.1083/jcb.153.3.599
中图分类号
Q2 [细胞生物学];
学科分类号
071009 ; 090102 ;
摘要
The translocation of fluorescently tagged PLC gamma and requirements for this process in cells stimulated with EGF were analyzed using real time fluorescence microscopy applied for the first time to monitor growth factor receptor-effector interactions. The translocation of PLC gamma to the plasma membrane required the functional Src homology 2 domains and was not affected by mutations in the pleckstrin homology domain or inhibition of phosphatidylinositol (PI) 3-kinase. An array of domains specific for PLC gamma isoforms was sufficient for this translocation. The dynamics of translocation to the plasma membrane and redistribution of PLC gamma, relative to localization of the EGF receptor and PI 4,5-biphosphate (PI 4,5-P-2), were shown. Colocalization with the receptor was observed in the plasma membrane and in membrane ruffles where PI 4,5-P-2 substrate could also be visualized. At later times, internalization of PLC gamma, which could lead to separation from the substrate, was observed. The data support a direct binding of PLC gamma to the receptor as the main site of the plasma membrane recruitment. The presence of PLC gamma in membrane structures and its access to the substrate appear to be transient and are followed by a rapid incorporation into intracellular vesicles, leading to downregulation of the PLC activity.
引用
收藏
页码:599 / 612
页数:14
相关论文
共 36 条
  • [1] COMPARTMENTALIZED SIGNAL-TRANSDUCTION BY RECEPTOR TYROSINE KINASES
    BAASS, PC
    DIGUGLIELMO, GM
    AUTHIER, F
    POSNER, BI
    BERGERON, JJM
    [J]. TRENDS IN CELL BIOLOGY, 1995, 5 (12) : 465 - 470
  • [2] Activation of phospholipase C-γ by phosphatidylinositol 3,4,5-trisphosphate
    Bae, YS
    Cantley, LG
    Chen, CS
    Kim, SR
    Kwon, KS
    Rhee, SG
    [J]. JOURNAL OF BIOLOGICAL CHEMISTRY, 1998, 273 (08) : 4465 - 4469
  • [3] Intracellular signal transduction: The role of endosomes
    Bevan, AP
    Drake, PG
    Bergeron, JJM
    Posner, BI
    [J]. TRENDS IN ENDOCRINOLOGY AND METABOLISM, 1996, 7 (01) : 13 - 21
  • [4] Endocytosis of functional epidermal growth factor receptor-green fluorescent protein chimera
    Carter, RE
    Sorkin, A
    [J]. JOURNAL OF BIOLOGICAL CHEMISTRY, 1998, 273 (52) : 35000 - 35007
  • [5] The role of individual SH2 domains in mediating association of phospholipase C-γ1 with the activated EGF receptor
    Chattopadhyay, A
    Vecchi, M
    Ji, QS
    Mernaugh, R
    Carpenter, G
    [J]. JOURNAL OF BIOLOGICAL CHEMISTRY, 1999, 274 (37) : 26091 - 26097
  • [6] PIP2 and PIP3: Complex roles at the cell surface
    Czech, MP
    [J]. CELL, 2000, 100 (06) : 603 - 606
  • [7] DeBell KE, 1999, MOL CELL BIOL, V19, P7388
  • [8] Activation of phospholipase Cγ by PI 3-kinase-induced PH domain-mediated membrane targeting
    Falasca, M
    Logan, SK
    Lehto, VP
    Baccante, G
    Lemmon, MA
    Schlessinger, J
    [J]. EMBO JOURNAL, 1998, 17 (02) : 414 - 422
  • [9] FUKAMI K, 1988, P NATL ACAD SCI USA, V85, P9067
  • [10] Effect of epidermal growth factor receptor internalization on regulation of the phospholipase C-γ1 signaling pathway
    Haugh, JM
    Schooler, K
    Wells, A
    Wiley, HS
    Lauffenburger, DA
    [J]. JOURNAL OF BIOLOGICAL CHEMISTRY, 1999, 274 (13) : 8958 - 8965