title: Cophosphorylation of amphiphysin I and dynamin I by Cdk5 regulates clathrin-mediated endocytosis of synaptic vesicles.
abstract: 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.
authors: Department of Physiology, Okayama University Graduate School of Medicine and Dentistry, Okayama 700-8558, Japan. tomikt@md.okayama-u.ac.jp
authors: Kazuhito
authors: K
authors: Tomizawa
authors: Satoshi
authors: S
authors: Sunada
authors: Yun-Fei
authors: YF
authors: Lu
authors: Yoshiya
authors: Y
authors: Oda
authors: Masahiro
authors: M
authors: Kinuta
authors: Toshio
authors: T
authors: Ohshima
authors: Taro
authors: T
authors: Saito
authors: Fan-Yan
authors: FY
authors: Wei
authors: Masayuki
authors: M
authors: Matsushita
authors: Sheng-Tian
authors: ST
authors: Li
authors: Kimiko
authors: K
authors: Tsutsui
authors: Shin-ichi
authors: S
authors: Hisanaga
authors: Katsuhiko
authors: K
authors: Mikoshiba
authors: Kohji
authors: K
authors: Takei
authors: Hideki
authors: H
authors: Matsui
mesh_terms: 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*
chemical_list: 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