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Abstract
While a significant amount is known about the biochemical signaling pathways of the Rho family GTPase Cdc42, a better understanding of how these signaling networks are coordinated in cells is required. In particular, the predominant subcellular sites where GTP-bound Cdc42 binds to its effectors, such as p21-activated kinase 1 (PAKI) and N-WASP, a homolog of the Wiskott-Aldritch syndrome protein, are still undetermined. Recent fluorescence resonance energy transfer (FRET) imaging experiments using activity biosensors show inconsistencies between the site of local activity of PAKI or N-WASP and the formation of specific membrane protrusion structures in the cell periphery. The data presented here demonstrate the localization of interactions by using multiphoton time-domain fluorescence lifetime imaging microscopy (FLIM). Our data here establish that activated Cdc42 interacts with PAKI in a nucleotide-dependent manner in the cell periphery, leading to Thr-423 phosphorylation of PAKI, particularly along the lengths of cell protrusion structures. In contrast, the majority of GFP-N-WASP undergoing FRET with Cy3-Cdc42 is localized within a transferrin receptor- and Rab11-positive endosomal compartment in breast carcinoma cells. These data reveal for the first time distinct spatial association patterns between Cdc42 and its key effector proteins controlling cytoskeletal remodeling.
Original language | English |
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Pages (from-to) | 1680-95 |
Number of pages | 16 |
Journal | Molecular and Cellular Biology |
Volume | 25 |
Issue number | 5 |
DOIs | |
Publication status | Published - 1 Mar 2005 |
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Dive into the research topics of 'Spatially distinct binding of Cdc42 to PAK1 and N/WASP in breast carcinoma cells'. Together they form a unique fingerprint.Projects
- 1 Finished
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Fellowship: Dr L Machesky - The Arp2/3 Complex in Actin- Based Motility.
Machesky, L.
1/10/00 → 30/09/05
Project: Research Councils