Visualization of spatially and temporally regulated N-WASP activity during cytoskeletal reorganization in living cells

Michael E. Ward, Jane Y. Wu*, Yi Rao

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

26 Scopus citations

Abstract

Members of the WASP/WAVE family of proteins are key regulators of cytoskeletal reorganization across a diverse range of cellular processes. Despite a wealth of biochemical data about WASP/WAVE regulation in vitro, our understanding of the in vivo regulation of these proteins is hampered by the inability to monitor subcellular regulation of their activities in living cells. Here we establish a fluorescence resonance energy transfer-based approach to visualize spatial and temporal regulation of neuronal Wiskott-Aldrich syndrome protein (N-WASP) activity in living cells. Using time-lapse microscopy, we characterize the activation of N-WASP in response to growth-factor stimulation, and we reveal that N-WASP is activated inside extending filopodia. Furthermore, we suggest a role of N-WASP in regulating membrane ruffling downstream of phosphatidylinositol 4,5-bisphosphate and Cdc42.

Original languageEnglish (US)
Pages (from-to)970-974
Number of pages5
JournalProceedings of the National Academy of Sciences of the United States of America
Volume101
Issue number4
DOIs
StatePublished - Jan 27 2004

ASJC Scopus subject areas

  • General

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