Cophosphorylation of amphiphysin I and dynamin I by Cdk5 regulates clathrin-mediated endocytosis of synaptic vesicles.
It has been thought that clathrin-mediated endocytosis is regulated by phosphorylation and dephosphorylation of many endocytic proteins, including amphiphysin I and dynamin I. Here, we show that Cdk5/p35-dependent cophosphorylation of amphiphysin I and dynamin I plays a critical role in such processes. Cdk5 inhibitors enhanced the electric stimulation-induced endocytosis in hippocampal neurons, and the endocytosis was also enhanced in the neurons of p35-deficient mice. Cdk5 phosphorylated the proline-rich domain of both amphiphysin I and dynamin I in vitro and in vivo. Cdk5-dependent phosphorylation of amphiphysin I inhibited the association with beta-adaptin. Furthermore, the phosphorylation of dynamin I blocked its binding to amphiphysin I. The phosphorylation of each protein reduced the copolymerization into a ring formation in a cell-free system. Moreover, the phosphorylation of both proteins completely disrupted the copolymerization into a ring formation. Finally, phosphorylation of both proteins was undetectable in p35-deficient mice.
Tomizawa
Kazuhito
K
Department of Physiology, Okayama University Graduate School of Medicine and Dentistry, Okayama 700-8558, Japan. tomikt@md.okayama-u.ac.jp
Sunada
Satoshi
S
Lu
Yun-Fei
YF
Oda
Yoshiya
Y
Kinuta
Masahiro
M
Ohshima
Toshio
T
Saito
Taro
T
Wei
Fan-Yan
FY
Matsushita
Masayuki
M
Li
Sheng-Tian
ST
Tsutsui
Kimiko
K
Hisanaga
Shin-ichi
S
Mikoshiba
Katsuhiko
K
Takei
Kohji
K
Matsui
Hideki
H
D033966:Adaptor Protein Complex beta Subunits / Q000235:genetics / Q000378:metabolism; D000818:Animals; D001370:Axonal Transport / Q000235:genetics; D002478:Cells, Cultured; D002966:Clathrin / Q000235:genetics / Q000378:metabolism; D051360:Cyclin-Dependent Kinase 5; D018844:Cyclin-Dependent Kinases / Q000378:metabolism*; D034284:Dynamin I / Q000235:genetics / Q000378:metabolism*; D004558:Electric Stimulation; D004705:Endocytosis / Q000187:drug effects / Q000235:genetics*; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005333:Fetus; D006624:Hippocampus / Q000201:enzymology; D046911:Macromolecular Substances; D051379:Mice; D018345:Mice, Knockout; D009419:Nerve Tissue Proteins / Q000172:deficiency / Q000235:genetics / Q000378:metabolism*; D010766:Phosphorylation / Q000187:drug effects; D011108:Polymers / Q000378:metabolism; D011485:Protein Binding / Q000187:drug effects / Q000235:genetics; D013572:Synaptic Vesicles / Q000201:enzymology*
D033966:Adaptor Protein Complex beta Subunits; D002966:Clathrin; D004791:Enzyme Inhibitors; D046911:Macromolecular Substances; D009419:Nerve Tissue Proteins; D011108:Polymers; C089495:neuronal Cdk5 activator (p25-p35); C075637:amphiphysin; D051360:Cyclin-Dependent Kinase 5; C495906:Cdk5 protein, mouse; D018844:Cyclin-Dependent Kinases; D034284:Dynamin I