Cophosphorylation of amphiphysin I and dynamin I by Cdk5 regulates clathrin-mediated endocytosis of synaptic vesicles

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

Research output: Contribution to journalArticle

153 Citations (Scopus)

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 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.

Original languageEnglish
Pages (from-to)813-824
Number of pages12
JournalJournal of Cell Biology
Volume163
Issue number4
DOIs
Publication statusPublished - 2003 Nov 24
Externally publishedYes

Fingerprint

Dynamin I
Clathrin
Synaptic Vesicles
Endocytosis
Phosphorylation
Proteins
Neurons
Cell-Free System
Proline
Electric Stimulation
amphiphysin

Keywords

  • Cyclin-dependent kinase
  • Endocytic protein
  • P35
  • Presynapse
  • Synaptosome

ASJC Scopus subject areas

  • Cell Biology

Cite this

Cophosphorylation of amphiphysin I and dynamin I by Cdk5 regulates clathrin-mediated endocytosis of synaptic vesicles. / Tomizawa, Kazuhito; Sunada, Satoshi; Lu, Yun Fei; Oda, Yoshiya; Kinuta, Masahiro; Ohshima, Toshio; Saito, Taro; Wei, Fan Yan; Matsushita, Masayuki; Li, Sheng Tian; Tsutsui, Kimiko; Hisanaga, Shin Ichi; Mikoshiba, Katsuhiko; Takei, Kohji; Matsui, Hideki.

In: Journal of Cell Biology, Vol. 163, No. 4, 24.11.2003, p. 813-824.

Research output: Contribution to journalArticle

Tomizawa, K, Sunada, S, Lu, YF, Oda, Y, Kinuta, M, Ohshima, T, Saito, T, Wei, FY, Matsushita, M, Li, ST, Tsutsui, K, Hisanaga, SI, Mikoshiba, K, Takei, K & Matsui, H 2003, 'Cophosphorylation of amphiphysin I and dynamin I by Cdk5 regulates clathrin-mediated endocytosis of synaptic vesicles', Journal of Cell Biology, vol. 163, no. 4, pp. 813-824. https://doi.org/10.1083/jcb.200308110
Tomizawa, Kazuhito ; Sunada, Satoshi ; Lu, Yun Fei ; Oda, Yoshiya ; Kinuta, Masahiro ; Ohshima, Toshio ; Saito, Taro ; Wei, Fan Yan ; Matsushita, Masayuki ; Li, Sheng Tian ; Tsutsui, Kimiko ; Hisanaga, Shin Ichi ; Mikoshiba, Katsuhiko ; Takei, Kohji ; Matsui, Hideki. / Cophosphorylation of amphiphysin I and dynamin I by Cdk5 regulates clathrin-mediated endocytosis of synaptic vesicles. In: Journal of Cell Biology. 2003 ; Vol. 163, No. 4. pp. 813-824.
@article{d20e4f842d0140feaf21b9a9dffff41f,
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 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.",
keywords = "Cyclin-dependent kinase, Endocytic protein, P35, Presynapse, Synaptosome",
author = "Kazuhito Tomizawa and Satoshi Sunada and Lu, {Yun Fei} and Yoshiya Oda and Masahiro Kinuta and Toshio Ohshima and Taro Saito and Wei, {Fan Yan} and Masayuki Matsushita and Li, {Sheng Tian} and Kimiko Tsutsui and Hisanaga, {Shin Ichi} and Katsuhiko Mikoshiba and Kohji Takei and Hideki Matsui",
year = "2003",
month = "11",
day = "24",
doi = "10.1083/jcb.200308110",
language = "English",
volume = "163",
pages = "813--824",
journal = "Journal of Cell Biology",
issn = "0021-9525",
publisher = "Rockefeller University Press",
number = "4",

}

TY - JOUR

T1 - Cophosphorylation of amphiphysin I and dynamin I by Cdk5 regulates clathrin-mediated endocytosis of synaptic vesicles

AU - Tomizawa, Kazuhito

AU - Sunada, Satoshi

AU - Lu, Yun Fei

AU - Oda, Yoshiya

AU - Kinuta, Masahiro

AU - Ohshima, Toshio

AU - Saito, Taro

AU - Wei, Fan Yan

AU - Matsushita, Masayuki

AU - Li, Sheng Tian

AU - Tsutsui, Kimiko

AU - Hisanaga, Shin Ichi

AU - Mikoshiba, Katsuhiko

AU - Takei, Kohji

AU - Matsui, Hideki

PY - 2003/11/24

Y1 - 2003/11/24

N2 - 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.

AB - 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.

KW - Cyclin-dependent kinase

KW - Endocytic protein

KW - P35

KW - Presynapse

KW - Synaptosome

UR - http://www.scopus.com/inward/record.url?scp=10744228914&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=10744228914&partnerID=8YFLogxK

U2 - 10.1083/jcb.200308110

DO - 10.1083/jcb.200308110

M3 - Article

C2 - 14623869

AN - SCOPUS:10744228914

VL - 163

SP - 813

EP - 824

JO - Journal of Cell Biology

JF - Journal of Cell Biology

SN - 0021-9525

IS - 4

ER -