This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Supplemental material
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrowReprints and Permissions
Right arrow Copyright Information
Right arrow Books from ASM Press
Right arrow MicrobeWorld
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Parsons, M.
Right arrow Articles by Ng, T.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Parsons, M.
Right arrow Articles by Ng, T.

 Previous Article  |  Next Article 

Molecular and Cellular Biology, March 2005, p. 1680-1695, Vol. 25, No. 5
0270-7306/05/$08.00+0     doi:10.1128/MCB.25.5.1680-1695.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Spatially Distinct Binding of Cdc42 to PAK1 and N-WASP in Breast Carcinoma Cells{dagger}

Maddy Parsons,1,{ddagger}* James Monypenny,2,{ddagger} Simon M. Ameer-Beg,3,{ddagger} Thomas H. Millard,4 Laura M. Machesky,4 Marion Peter,1 Melanie D. Keppler,1 Giampietro Schiavo,5 Rose Watson,6 Jonathan Chernoff,7 Daniel Zicha,2 Borivoj Vojnovic,3 and Tony Ng1

Randall Centre, King's College London, Guy's Medical School Campus,1 Light Microscopy Laboratory,2 Molecular NeuroPathoBiology Laboratory,5 Electron Microscopy Unit,6 Cancer Research UK London Research Institute, London, Gray Cancer Institute, Mount Vernon Hospital, Northwood, Middlesex,3 School of Biosciences, Division of Molecular and Cell Biology, University of Birmingham, Birmingham United Kingdom,4 Fox Chase Cancer Center, Philadelphia, Pennsylvania7

Received 3 June 2004/ Returned for modification 12 August 2004/ Accepted 26 November 2004

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 (PAK1) 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 PAK1 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 PAK1 in a nucleotide-dependent manner in the cell periphery, leading to Thr-423 phosphorylation of PAK1, 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.

* Corresponding author. Mailing address: Randall Centre, King's College London, 3rd Floor, New Hunt's House, Guy's Medical School Campus, London SE1 1UL, United Kingdom. Phone: 44 (0) 20 7848 6835. Fax: 44 (0) 20 7848 6435. E-mail: maddy.parsons{at}kcl.ac.uk.

{dagger} Supplemental material for this article may be found at http://mcb.asm.org/.

{ddagger} M.P., J.M., and S.M.A.-B. contributed equally to this study.

Molecular and Cellular Biology, March 2005, p. 1680-1695, Vol. 25, No. 5
0022-538X/05/$08.00+0     doi:10.1128/MCB.25.5.1680-1695.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Dharmalingam, E., Haeckel, A., Pinyol, R., Schwintzer, L., Koch, D., Kessels, M. M., Qualmann, B. (2009). F-BAR Proteins of the Syndapin Family Shape the Plasma Membrane and Are Crucial for Neuromorphogenesis. J. Neurosci. 29: 13315-13327 [Abstract] [Full Text]  
  • Landry, M.-C., Sicotte, A., Champagne, C., Lavoie, J. N. (2009). Regulation of Cell Death by Recycling Endosomes and Golgi Membrane Dynamics via a Pathway Involving Src-family kinases, Cdc42 and Rab11a. Mol. Biol. Cell 20: 4091-4106 [Abstract] [Full Text]  
  • Makrogianneli, K., Carlin, L. M., Keppler, M. D., Matthews, D. R., Ofo, E., Coolen, A., Ameer-Beg, S. M., Barber, P. R., Vojnovic, B., Ng, T. (2009). Integrating Receptor Signal Inputs That Influence Small Rho GTPase Activation Dynamics at the Immunological Synapse. Mol. Cell. Biol. 29: 2997-3006 [Abstract] [Full Text]  
  • Killock, D. J., Parsons, M., Zarrouk, M., Ameer-Beg, S. M., Ridley, A. J., Haskard, D. O., Zvelebil, M., Ivetic, A. (2009). In Vitro and in Vivo Characterization of Molecular Interactions between Calmodulin, Ezrin/Radixin/Moesin, and L-selectin. J. Biol. Chem. 284: 8833-8845 [Abstract] [Full Text]  
  • Barber, P.R, Ameer-Beg, S.M, Gilbey, J, Carlin, L.M, Keppler, M, Ng, T.C, Vojnovic, B (2009). Multiphoton time-domain fluorescence lifetime imaging microscopy: practical application to protein-protein interactions using global analysis. J R Soc Interface 6: S93-S105 [Abstract] [Full Text]  
  • Parsons, M., Adams, J. C. (2008). Rac regulates the interaction of fascin with protein kinase C in cell migration. J. Cell Sci. 121: 2805-2813 [Abstract] [Full Text]  
  • Rodal, A. A., Motola-Barnes, R. N., Littleton, J. T. (2008). Nervous Wreck and Cdc42 Cooperate to Regulate Endocytic Actin Assembly during Synaptic Growth. J. Neurosci. 28: 8316-8325 [Abstract] [Full Text]  
  • Parsons, M., Messent, A. J., Humphries, J. D., Deakin, N. O., Humphries, M. J. (2008). Quantification of integrin receptor agonism by fluorescence lifetime imaging. J. Cell Sci. 121: 265-271 [Abstract] [Full Text]  
  • Prag, S., Parsons, M., Keppler, M. D., Ameer-Beg, S. M., Barber, P., Hunt, J., Beavil, A. J., Calvert, R., Arpin, M., Vojnovic, B., Ng, T. (2007). Activated Ezrin Promotes Cell Migration through Recruitment of the GEF Dbl to Lipid Rafts and Preferential Downstream Activation of Cdc42. Mol. Biol. Cell 18: 2935-2948 [Abstract] [Full Text]  
  • Panner, A., Nakamura, J. L., Parsa, A. T., Rodriguez-Viciana, P., Berger, M. S., Stokoe, D., Pieper, R. O. (2006). mTOR-Independent Translational Control of the Extrinsic Cell Death Pathway by RalA. Mol. Cell. Biol. 26: 7345-7357 [Abstract] [Full Text]  


Copyright © 2009 by the American Society for Microbiology.   For an alternate route to Journals.ASM.org, visit: http://intl-journals.asm.org | More Info»