UV induces tyrosine kinase-independent internalisation and endosome arrest of the EGF receptor.
We have compared the activation and trafficking of epidermal growth factor receptor (EGFR) induced by UV light and EGF. Tyrosine phosphorylation of EGFR was not detected in UV-exposed cells by immunoblotting of whole cell lysates or EGFR immunoprecipitates with antibodies specific for each of the five activated autophosphorylation sites of EGFR. In addition, EGFR of UV-irradiated cells did not demonstrate increased (32)P-incorporation. However, UV-exposed cells demonstrated a gel mobility shift of EGFR, which was not abolished by alkaline phosphatase treatment. UV-exposure did not induce dimerisation of EGFR. Furthermore, UV induced internalisation of EGFR without polyubiquitination or degradation. UV-exposed EGFR was transferred to early endosomes and arrested in transferrin-accessible endosomes close to the cell surface. Whereas inhibition of the EGFR tyrosine kinase effectively inhibited tyrosine phosphorylation and internalisation of EGF-activated EGFR, internalisation of UV-exposed EGFR was unaffected. UV induced neither relocalisation of Shc and Grb2 nor activation of Raf, but activation of MEK and MAPK was observed. Our work indicates that UV induces internalisation of EGFR independent of its phosphorylation or receptor tyrosine kinase activation, and altered EGFR trafficking compared with ligand-activated receptor. In addition, MAPK activation by UV does not appear to be mediated by EGFR activation.
Oksvold
Morten P
MP
Laboratory for Toxicopathology, Institute of Pathology, The National Hospital, University of Oslo, N-0027 Oslo, Norway. m.p.oksvold@labmed.uio.no
Huitfeldt
Henrik S
HS
Østvold
Anne Carine
AC
Skarpen
Ellen
E
D000469:Alkaline Phosphatase / Q000378:metabolism; D004705:Endocytosis / Q000528:radiation effects*; D011992:Endosomes / Q000378:metabolism*; D004789:Enzyme Activation; D066246:ErbB Receptors / Q000378:metabolism* / Q000502:physiology; D006367:HeLa Cells; D006801:Humans; D010766:Phosphorylation; D021381:Protein Transport; D015398:Signal Transduction / Q000528:radiation effects; D014466:Ultraviolet Rays
D066246:ErbB Receptors; D000469:Alkaline Phosphatase