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MBC in Press, published online ahead of print February 20, 2004
Mol. Biol. Cell 10.1091/mbc.E03-07-0493

A more recent version of this article appeared on May 1, 2004
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Submitted on July 15, 2003
Revised on February 1, 2004
Accepted on February 2, 2004

Regulation of dendritic branching and filopodia formation in hippocampal neurons by specific acylated protein motifs

Catherine Gauthier-Campbell1, David S. Bredt2, Timothy H. Murphy1, and Alaa El-Din El-Husseini1*

1 Department of Psychiatry and the Brain Research Centre, 2255 Wesbrook Mall, University of British Columbia, Vancouver, BC, Canada, V6T 1Z3
2 Department of Physiology, University of California at San Francisco, San Francisco, CA 94143-0444

* Corresponding author. E-mail address: alaa{at}interchange.ubc.ca.

Although neuronal axons and dendrites with their associated filopodia and spines exhibit a profound cell polarity, the mechanism by which they develop is largely unknown. Here, we demonstrate that specific palmitoylated protein motifs, characterized by two adjacent cysteines and nearby basic residues, are sufficient to induce filopodial extensions in heterologous cells and to increase the number of filopodia and the branching of dendrites and axons in neurons. Such motifs are present at the N-terminus of GAP-43 and the C-terminus of paralemmin, two neuronal proteins implicated in cytoskeletal organization and filopodial outgrowth. Filopodia induction is blocked by mutations of the palmitoylated sites or by treatment with 2-bromopalmitate, an agent that inhibits protein palmitoylation. Moreover, overexpression of a constitutively active form of ARF-6, a GTPase that regulates membrane cycling and dendritic branching reversed the effects of the acylated protein motifs. Filopodia induction by the specific palmitoylated motifs was also reduced upon overexpression of a dominant negative form of the GTPase cdc42. These results demonstrate that select dually lipidated protein motifs trigger changes in the development and growth of neuronal processes.




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