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 β-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.
Cophosphorylation of amphiphysin I and dynamin I by Cdk5 regulates clathrin-mediated endocytosis of synaptic vesicles
The online version of this article includes supplemental material.
Abbreviations used in this paper: DLS, dynamic light scattering; ESI, electrospray ionization; LC, liquid chromatography; MALDI-MS, matrix assisted laser desorption/ionization mass spectrometry; PRD, proline-rich domain; SH3, Src homology 3; VDCC, voltage-dependent calcium channel.
Kazuhito Tomizawa, Satoshi Sunada, Yun-Fei Lu, Yoshiya Oda, Masahiro Kinuta, Toshio Ohshima, Taro Saito, Fan-Yan Wei, Masayuki Matsushita, Sheng-Tian Li, Kimiko Tsutsui, Shin-ichi Hisanaga, Katsuhiko Mikoshiba, Kohji Takei, Hideki Matsui; Cophosphorylation of amphiphysin I and dynamin I by Cdk5 regulates clathrin-mediated endocytosis of synaptic vesicles . J Cell Biol 24 November 2003; 163 (4): 813–824. doi: https://doi.org/10.1083/jcb.200308110
Download citation file:
Sign in
Client Account
Sign in via your Institution
Sign in via your InstitutionEmail alerts
Advertisement
Advertisement