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Originally published as MBC in Press, 10.1091/mbc.E08-08-0890 on October 15, 2008

Vol. 19, Issue 12, 5347-5359, December 2008

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Isoform and Splice-Variant Specific Functions of Dynamin-2 Revealed by Analysis of Conditional Knock-Out Cells

Ya-Wen Liu, Mark C. Surka, Thomas Schroeter, Vasyl Lukiyanchuk, and Sandra L. Schmid

Department of Cell Biology, The Scripps Research Institute, La Jolla, CA 92037

Submitted August 29, 2008; Revised October 1, 2008; Accepted October 3, 2008
Monitoring Editor: David G. Drubin

InCytes from MBC

Dynamin (Dyn) is a multifunctional GTPase implicated in several cellular events, including endocytosis, intracellular trafficking, cell signaling, and cytokinesis. The mammalian genome encodes three isoforms, Dyn1, Dyn2, and Dyn3, and several splice variants of each, leading to the suggestion that distinct isoforms and/or distinct splice variants might mediate distinct cellular functions. We generated a conditional Dyn2 KO cell line and performed knockout and reconstitution experiments to explore the isoform- and splice variant specific cellular functions of ubiquitously expressed Dyn2. We find that Dyn2 is required for clathrin-mediated endocytosis (CME), p75 export from the Golgi, and PDGF-stimulated macropinocytosis and cytokinesis, but not for other endocytic pathways. Surprisingly, CME and p75 exocytosis were efficiently rescued by reintroduction of Dyn2, but not Dyn1, suggesting that these two isoforms function differentially in vesicular trafficking in nonneuronal cells. Both isoforms rescued macropinocytosis and cytokinesis, suggesting that dynamin function in these processes might be mechanistically distinct from its role in CME. Although all four Dyn2 splice variants could equally restore CME, Dyn2ba and -bb were more effective at restoring p75 exocytosis. This splice variant specificity correlated with their differential targeting to the Golgi. These studies reveal isoform and splice-variant specific functions for Dyn2.

This was published online ahead of print in MBC in Press (http://www.molbiolcell.org/cgi/doi/10.1091/mbc.E08-08-0890) on October 15, 2008.

Address correspondence to: Sandra L. Schmid (slschmid{at}scripps.edu)


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InCytes from MBC, December 2008

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