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MBC in Press, published online ahead of print November 19, 2008
Mol. Biol. Cell 10.1091/mbc.E08-08-0891

A more recent version of this article appeared on January 1, 2009 Originally published as MBC in Press, 10.1091/mbc.E08-08-0891 on November 5, 2008
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Submitted on September 2, 2008
Revised on October 21, 2008
Accepted on October 29, 2008

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

Susana B. Salvarezza,* Sylvie Deborde,* Ryan Schreiner,* Fabien Campagne,{dagger} Michael M. Kessels,{ddagger} Britta Qualmann,{ddagger} Alfredo Caceres,{sect} 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; {sect}Instituto de Investigacion Medica Mercedes y Martin Ferreyra, 5000 Cordoba, Argentina.; {ddagger}Institute for Biochemistry I, Friedrich Schiller University Jena, 07743 Jena, Germany

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 MDCK 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 siRNA, 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-GFP but did not interfere with the apical PM marker, GPI-GFP, or the basolateral PM marker, NCAM-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.

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






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