Regulation of amphiphysin1 by mitogen-activated protein kinase: its significance in nerve growth factor receptor-mediated endocytosis.
Amphiphysin1, which can simultaneously bind to dynamin1 and the clathrin adaptor AP-2, is essential for dynamin1 recruitment during receptor-mediated endocytosis, but little is known about its regulatory mechanism. Here, we purified a 120-kDa mitogen-activated protein kinase (MAPK) substrate protein from porcine brains and identified the protein as amphiphysin1. Serine phosphorylation of amphiphysin1 was rapidly induced by nerve growth factor (NGF) in PC12 cells, and the induction was blocked by a MAPK inhibitor. Furthermore, when phosphorylated by MAPK in vitro or by NGF treatment in vivo, amphiphysin1 failed to bind to AP-2, but its association with dynamin1 was unaffected. Consistent with this, mutation of consensus MAPK phosphorylation sites increased amphiphysin1 binding to AP-2 and their intracellular colocalization. Thus, we propose that MAPK phosphorylation of amphiphysin1 controls NGF receptor/TrkA-mediated endocytosis by terminating the amphiphysin1-AP-2 interaction. This perhaps helps to regulate the availability of amphiphysin1-dynamin1 complexes for binding to the endocytic vesicle.
Shang
Wei Hao
WH
Department of Molecular Biology and Biochemistry, Shinshu University School of Medicine, Matsumoto, Nagano, Japan.
Adachi
Yoshifumi
Y
Nakamura
Akihiro
A
Copeland
Terry
T
Kim
Seung Ryul
SR
Kamata
Tohru
T
D033962:Adaptor Protein Complex 2 / Q000378:metabolism; D000818:Animals; D004591:Electrophoresis, Polyacrylamide Gel; D004705:Endocytosis; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005419:Flavonoids / Q000494:pharmacology; D005786:Gene Expression Regulation; D005982:Glutathione Transferase / Q000378:metabolism; D015151:Immunoblotting; D007150:Immunohistochemistry; D020935:MAP Kinase Signaling System / Q000502:physiology*; D008856:Microscopy, Fluorescence; D008954:Models, Biological; D009154:Mutation; D020932:Nerve Growth Factor / Q000378:metabolism*; D009419:Nerve Tissue Proteins / Q000096:biosynthesis* / Q000378:metabolism; D016716:PC12 Cells; D019606:Phosphoamino Acids / Q000378:metabolism; D010766:Phosphorylation; D011233:Precipitin Tests; D011485:Protein Binding; D051381:Rats; D013347:Subcellular Fractions; D013552:Swine; D014162:Transfection
D033962:Adaptor Protein Complex 2; D004791:Enzyme Inhibitors; D005419:Flavonoids; D009419:Nerve Tissue Proteins; D019606:Phosphoamino Acids; C075637:amphiphysin; D020932:Nerve Growth Factor; D005982:Glutathione Transferase; C093973:2-(2-amino-3-methoxyphenyl)-4H-1-benzopyran-4-one