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
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