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Originally published as MBC in Press, 10.1091/mbc.E08-08-0891 on November 19, 2008 Originally published as MBC in Press, 10.1091/mbc.E08-08-0891 on November 5, 2008

Vol. 20, Issue 1, 438-451, January 1, 2009

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LIM Kinase 1 and Cofilin Regulate Actin Filament Population Required for Dynamin-dependent Apical Carrier Fission from the Trans-Golgi Network

Susana B. Salvarezza*, Sylvie Deborde*, Ryan Schreiner*, Fabien Campagne{dagger}, Michael M. Kessels{ddagger}, Britta Qualmann{ddagger}, Alfredo Caceres§, Geri Kreitzer||, and Enrique Rodriguez-Boulan*,||

*Margaret Dyson Vision Research Institute and ||Department of Cell and Developmental Biology and {dagger}Department of Physiology and Biophysics, HRH Prince Alwaleed Bin Talal Bin Abdulaziz Alsaud Institute for Computational Biomedicine, Weill Medical College of Cornell University, New York, NY 10065; §Instituto de Investigacion Medica Mercedes y Martin Ferreyra, 5000 Cordoba, Argentina; and {ddagger}Institute for Biochemistry I, Friedrich Schiller University Jena, 07743 Jena, Germany

Submitted September 2, 2008; Revised October 21, 2008; Accepted October 29, 2008
Monitoring Editor: Sandra L. Schmid

The functions of the actin cytoskeleton in post-Golgi trafficking are still poorly understood. Here, we report the role of LIM Kinase 1 (LIMK1) and its substrate cofilin in the trafficking of apical and basolateral proteins in Madin-Darby canine kidney cells. Our data indicate that LIMK1 and cofilin organize a specialized population of actin filaments at the Golgi complex that is selectively required for the emergence of an apical cargo route to the plasma membrane (PM). Quantitative pulse-chase live imaging experiments showed that overexpression of kinase-dead LIMK1 (LIMK1-KD), or of LIMK1 small interfering RNA, or of an activated cofilin mutant (cofilin S3A), selectively slowed down the exit from the trans-Golgi network (TGN) of the apical PM marker p75-green fluorescent protein (GFP) but did not interfere with the apical PM marker glycosyl phosphatidylinositol-YFP or the basolateral PM marker neural cell adhesion molecule-GFP. High-resolution live imaging experiments of carrier formation and release by the TGN and analysis of peri-Golgi actin dynamics using photoactivatable GFP suggest a scenario in which TGN-localized LIMK1-cofilin regulate a population of actin filaments required for dynamin-syndapin-cortactin–dependent generation and/or fission of precursors to p75 transporters.

This was published online ahead of print in MBC in Press (http://www.molbiolcell.org/cgi/doi/10.1091/mbc.E08-08-0891) on November 5, 2008.

Address correspondence to: Enrique Rodriguez-Boulan (boulan{at}med.cornell.edu)






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