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Published online January 21, 2008
doi:10.1083/jcb.200710039
The Journal of Cell Biology, Vol. 180, No. 2, 295-303
The Rockefeller University Press, 0021-9525 $30.00
© 2008 Spiliotis et al.
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 Epithelial polarity requires septin coupling of vesicle transport to polyglutamylated microtubules

Elias T. Spiliotis1, Stephen J. Hunt1, Qicong Hu1, Makoto Kinoshita2, and W. James Nelson1

1 Departments of Biology, and Molecular and Cellular Physiology, Stanford University, Stanford, CA 94305
2 Biochemistry and Cell Biology Unit, HMRO, Kyoto University, Kyoto 606-8501, Japan

Correspondence to Elias T. Spiliotis: eliass{at}stanford.edu; or W. James Nelson: wjnelson{at}stanford.edu

In epithelial cells, polarized growth and maintenance of apical and basolateral plasma membrane domains depend on protein sorting from the trans-Golgi network (TGN) and vesicle delivery to the plasma membrane. Septins are filamentous GTPases required for polarized membrane growth in budding yeast, but whether they function in epithelial polarity is unknown. Here, we show that in epithelial cells septin 2 (SEPT2) fibers colocalize with a subset of microtubule tracks composed of polyglutamylated (polyGlu) tubulin, and that vesicles containing apical or basolateral proteins exit the TGN along these SEPT2/polyGlu microtubule tracks. Tubulin-associated SEPT2 facilitates vesicle transport by maintaining polyGlu microtubule tracks and impeding tubulin binding of microtubule-associated protein 4 (MAP4). Significantly, this regulatory step is required for polarized, columnar-shaped epithelia biogenesis; upon SEPT2 depletion, cells become short and fibroblast-shaped due to intracellular accumulation of apical and basolateral membrane proteins, and loss of vertically oriented polyGlu microtubules. We suggest that septin coupling of the microtubule cytoskeleton to post-Golgi vesicle transport is required for the morphogenesis of polarized epithelia.

S. Hunt and Q. Hu contributed equally to this paper.

Abbreviations used in this paper: C-MBD-MAP4, C-terminal microtubule binding domain of microtubule-associated protein 4; GalTase, galactosyltransferase; gpi, glycosyl phosphatidylinositol; LDL-R, low density lipoprotein receptor; MAP, microtubule-associated protein; p75, neurotrophin receptor protein 75; PKD, protein kinase D1; polyGlu, polyglutamylated; VSV-G, vesicular stomatitis virus G protein.


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