Two domains of the epidermal growth factor receptor are involved in cytoskeletal interactions.
Epidermal growth factor receptor can interact directly with F-actin through an actin-binding domain. In the present study, a mutant EGFR, lacking a previously identified actin-binding domain (ABD 1), was still able to bind elements of the cytoskeleton. A second EGFR actin-binding domain (ABD 2) was identified in the region of the receptor that includes Tyr-1148 by a yeast two-hybrid assay. GST fusion proteins comprising ABD 1 or ABD 2 bound actin in vitro and competed for actin-binding with the full-length EGFR. EGFR binding to actin was also studied in intact cells using fluorescence resonance energy transfer (FRET). The localization of the EGFR/actin-binding complex changed after EGF stimulation. Fusion proteins containing mutations in ABD1 or ABD2 did not display a FRET signal. The results lead to the conclusion that the interaction between ABD1 and ABD2 and actin during EGF-induced signal transduction, and thus between EGFR and actin, are important in cell activation.
Song
Wei
W
Key Laboratory of Molecular Cell Biology, Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai, China.
Wu
Jing
J
Ge
Gaoxiang
G
Lin
Qishui
Q
D000199:Actins / Q000032:analysis / Q000378:metabolism*; D000818:Animals; D001667:Binding, Competitive; D019556:COS Cells; D002522:Chlorocebus aethiops; D003599:Cytoskeleton / Q000378:metabolism*; D066246:ErbB Receptors / Q000032:analysis / Q000235:genetics / Q000378:metabolism*; D031541:Fluorescence Resonance Energy Transfer; D005982:Glutathione Transferase / Q000737:chemistry / Q000235:genetics; D006801:Humans; D008247:Lysosomes / Q000737:chemistry / Q000378:metabolism; D009154:Mutation; D017434:Protein Structure, Tertiary / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000032:analysis / Q000235:genetics / Q000378:metabolism; D020798:Two-Hybrid System Techniques
D000199:Actins; D011993:Recombinant Fusion Proteins; D005982:Glutathione Transferase; D066246:ErbB Receptors