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First published online 30 September 2008
doi: 10.1242/jcs.032847

Journal of Cell Science 121, 3413-3421 (2008)
Published by The Company of Biologists 2008
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Research Article

Murine CENPF interacts with syntaxin 4 in the regulation of vesicular transport

Ryan D. Pooley1,*, Katherine L. Moynihan1,*, Victor Soukoulis1, Samyukta Reddy1, Richard Francis2, Cecilia Lo2, Li-Jun Ma3 and David M. Bader1,{ddagger}

1 Stahlman Cardiovascular Research Laboratories, Program for Developmental Biology, and Department of Medicine, Vanderbilt University Medical Center, Nashville, TN 37232-6300, USA
2 Laboratory for Developmental Biology, NIHLBI, Bethesda, MD 20892-1583, USA
3 Department of Pathology, Vanderbilt University Medical Center, Nashville, TN 37232-2561, USA

{ddagger} Author for correspondence (e-mail: david.bader{at}vanderbilt.edu)

Accepted 21 July 2008

Syntaxin 4 is a component of the SNARE complex that regulates membrane docking and fusion. Using a yeast two-hybrid screen, we identify a novel interaction between syntaxin 4 and cytoplasmic murine CENPF, a protein previously demonstrated to associate with the microtubule network and SNAP-25. The binding domain for syntaxin 4 in CENPF was defined by yeast two-hybrid assay and co-immunoprecipitation. Confocal analyses in cell culture reveal a high degree of colocalization between endogenously expressed proteins in interphase cells. Additionally, the endogenous SNARE proteins can be isolated as a complex with CENPF in immunoprecipitation experiments. Further analyses demonstrate that murine CENPF and syntaxin 4 colocalize with components of plasma membrane recycling: SNAP-25 and VAMP2. Depletion of endogenous CENPF disrupts GLUT4 trafficking whereas expression of a dominant-negative form of CENPF inhibits cell coupling. Taken together, these studies demonstrate that CENPF provides a direct link between proteins of the SNARE system and the microtubule network and indicate a diverse role for murine CENPF in vesicular transport.

Key words: CENPF, Syntaxin 4, GLUT4, Rab11a, VAMP2, SNAP-25






© The Company of Biologists Ltd 2008