Src-dependent tyrosine phosphorylation regulates dynamin self-assembly and ligand-induced endocytosis of the epidermal growth factor receptor.
Endocytosis of ligand-activated receptors requires dynamin-mediated GTP hydrolysis, which is regulated by dynamin self-assembly. Here, we demonstrate that phosphorylation of dynamin I by c-Src induces its self-assembly and increases its GTPase activity. Electron microscopic analyses reveal that tyrosine-phosphorylated dynamin I spontaneously self-assembles into large stacks of rings. Tyrosine 597 was identified as being phosphorylated both in vitro and in cultured cells following epidermal growth factor receptor stimulation. The replacement of tyrosine 597 with phenylalanine impairs Src kinase-induced dynamin I self-assembly and GTPase activity in vitro. Expression of Y597F dynamin I in cells attenuates agonist-driven epidermal growth factor receptor internalization. Thus, c-Src-mediated tyrosine phosphorylation is required for the function of dynamin in ligand-induced signaling receptor internalization.
Ahn
Seungkirl
S
Howard Hughes Medical Institute, Department of Pharmacology, Duke University Medical Center, Durham, North Carolina 27710, USA.
Kim
Jihee
J
Lucaveche
Carmen L
CL
Reedy
Mary C
MC
Luttrell
Louis M
LM
Lefkowitz
Robert J
RJ
Daaka
Yehia
Y
D000818:Animals; D019556:COS Cells; D000081247:CSK Tyrosine-Protein Kinase; D034284:Dynamin I; D034281:Dynamins; D004705:Endocytosis; D066246:ErbB Receptors / Q000378:metabolism*; D020558:GTP Phosphohydrolases / Q000378:metabolism*; D008024:Ligands; D010766:Phosphorylation; D011487:Protein Conformation; D011505:Protein-Tyrosine Kinases / Q000378:metabolism; D051381:Rats; D014162:Transfection; D019061:src-Family Kinases / Q000378:metabolism*
D008024:Ligands; D066246:ErbB Receptors; D011505:Protein-Tyrosine Kinases; D000081247:CSK Tyrosine-Protein Kinase; D019061:src-Family Kinases; D034284:Dynamin I; D020558:GTP Phosphohydrolases; D034281:Dynamins