Neuronal IP3 3-Kinase is an F-actin-bundling Protein: Role in Dendritic Targeting and Regulation of Spine Morphology

被引:114
作者
Johnson, Hong W. [1 ]
Schell, Michael J. [1 ]
机构
[1] Uniformed Serv Univ Hlth Sci, Dept Pharmacol, Bethesda, MD 20814 USA
关键词
INOSITOL 1,4,5-TRISPHOSPHATE 3-KINASE; RAT-BRAIN; CALCIUM; CELLS; LOCALIZATION; PLASTICITY; 1,3,4,5-TETRAKISPHOSPHATE; OVEREXPRESSION; REORGANIZATION; 5-PHOSPHATASE;
D O I
10.1091/mbc.E09-01-0083
中图分类号
Q2 [细胞生物学];
学科分类号
071009 ; 090102 ;
摘要
The actin microstructure in dendritic spines is involved in synaptic plasticity. Inositol trisphosphate 3-kinase A (ITPKA) terminates Ins(1,4,5)P-3 signals emanating from spines and also binds filamentous actin (F-actin) through its amino terminal region (amino acids 1-66, N66). Here we investigated how ITPKA, independent of its kinase activity, regulates dendritic spine F-actin microstructure. We show that the N66 region of the protein mediates F-actin bundling. An N66 fusion protein bundled F-actin in vitro, and the bundling involved N66 dimerization. By mutagenesis we identified a point mutation in a predicted helical region that eliminated both F-actin binding and bundling, rendering the enzyme cytosolic. A fusion protein containing a minimal helical region (amino acids 9-52, N9-52) bound F-actin in vitro and in cells, but had lower affinity. In hippocampal neurons, GFP-tagged N66 expression was highly polarized, with targeting of the enzyme predominantly to spines. By contrast, N9-52-GFP expression occurred in actin-rich structures in dendrites and growth cones. Expression of N66-GFP tripled the length of dendritic protrusions, induced longer dendritic spine necks, and induced polarized actin motility in time-lapse assays. These results suggest that, in addition to its ability to regulate intracellular Ca2+ via Ins(1,4,5)P-3 metabolism, ITPKA regulates structural plasticity.
引用
收藏
页码:5166 / 5180
页数:15
相关论文
共 52 条
[31]   A myosin motor that selects bundled actin for motility [J].
Nagy, Stanislav ;
Ricca, Benjamin L. ;
Norstrom, Melanie F. ;
Courson, David S. ;
Brawley, Crista M. ;
Smithback, Philip A. ;
Rock, Ronald S. .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 2008, 105 (28) :9616-9620
[32]   The nature of the globular-to fibrous-actin transition [J].
Oda, Toshiro ;
Iwasa, Mitsusada ;
Aihara, Tomoki ;
Maeda, Yuichiro ;
Narita, Akihiro .
NATURE, 2009, 457 (7228) :441-445
[33]   The role of CaMKII as an F-actin-bundling protein crucial for maintenance of dendritic spine structure [J].
Okamoto, Ken-Ichi ;
Narayanan, Radhakrishnan ;
Lee, Sang H. ;
Murata, Kazuyoshi ;
Hayashi, Yasunori .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 2007, 104 (15) :6418-6423
[34]   A polybasic motif allows N-WASP to act as a sensor of PIP2 density [J].
Papayannopoulos, V ;
Co, C ;
Prehoda, KE ;
Snapper, S ;
Taunton, J ;
Lim, WA .
MOLECULAR CELL, 2005, 17 (02) :181-191
[35]   Ins(1,4,5)P3 metabolism and the family of IP3-3Kinases [J].
Pattni, K ;
Banting, G .
CELLULAR SIGNALLING, 2004, 16 (06) :643-654
[36]   Inositol polyphosphate multikinase: metabolic architect of nuclear inositides [J].
Resnick, Adam Cain ;
Saiardi, Adolfo .
FRONTIERS IN BIOSCIENCE-LANDMARK, 2008, 13 :856-866
[37]   Lifeact: a versatile marker to visualize F-actin [J].
Riedl, Julia ;
Crevenna, Alvaro H. ;
Kessenbrock, Kai ;
Yu, Jerry Haochen ;
Neukirchen, Dorothee ;
Bista, Michal ;
Bradke, Frank ;
Jenne, Dieter ;
Holak, Tad A. ;
Werb, Zena ;
Sixt, Michael ;
Wedlich-Soldner, Roland .
NATURE METHODS, 2008, 5 (07) :605-607
[38]   Synaptically activated Ca2+ release from internal stores in CNS neurons [J].
Ross, WN ;
Nakamura, T ;
Watanabe, S ;
Larkum, M ;
Lasser-Ross, N .
CELLULAR AND MOLECULAR NEUROBIOLOGY, 2005, 25 (02) :283-295
[39]   Analysis of synaptic ultrastructure without fixative using high-pressure freezing and tomography [J].
Rostaing, Philippe ;
Real, Eleonore ;
Siksou, Lea ;
Lechaire, Jean-Pierre ;
Boudier, Thomas ;
Boeckers, Tobias M. ;
Gertler, Frank ;
Gundelfinger, Eckart D. ;
Triller, Antoine ;
Marty, Serge .
EUROPEAN JOURNAL OF NEUROSCIENCE, 2006, 24 (12) :3463-3474
[40]   Calcium-triggered exit of F-actin and IP3 3-kinase A from dendritic spines is rapid and reversible [J].
Schell, Michael J. ;
Irvine, Robin F. .
EUROPEAN JOURNAL OF NEUROSCIENCE, 2006, 24 (09) :2491-2503