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MBC in Press, published online ahead of print March 23, 2005
Mol. Biol. Cell 10.1091/mbc.E04-11-1001

A more recent version of this article appeared on June 1, 2005
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Submitted on November 16, 2004
Revised on February 7, 2005
Accepted on March 14, 2005

Myosin Va Transports Dense Core Secretory Vesicles in Pancreatic MIN6 {beta}-Cells

Aniko Varadi,*{dagger} Takashi Tsuboi,* and Guy A. Rutter*

*Henry Wellcome Laboratories for Integrated Cell Signalling and Department of Biochemistry, School of Medical Sciences, University of Bristol, Bristol BS8 1TD, United Kingdom; {dagger}Genomics Research Institute, Centre for Research in Biomedicine, University of the West of England, Frenchay Campus, Bristol BS16 1QY, United Kingdom

Monitoring Editor: Benjamin Glick

The role of unconventional myosins in neuroendocrine cells is not fully understood, with involvement suggested in the movement of both secretory vesicles and mitochondria. Here, we demonstrate colocalisation of myosin Va (MyoVa) with insulin in pancreatic {beta}-cells and show that MyoVa copurifies with insulin in density gradients and with the vesicle marker phogrin-EGFP upon fluorescence-activated sorting of vesicles. By contrast, MyoVa immunoreactivity was poorly colocalized with mitochondrial or other markers. Demonstrating an important role for MyoVa in the recruitment of secretory vesicles to the cell surface, a reduction of MyoVa protein levels achieved by RNA interference caused a significant decrease in glucose- or depolarisation-stimulated insulin secretion. Similarly, expression of the dominant-negative-acting globular tail domain of MyoVa decreased by ~50% the number of vesicles docked at the plasma membrane and by 87% the number of depolarisation-stimulated exocytotic events detected by total internal reflection fluorescence microscopy. We conclude that MyoVa-driven movements of vesicles along the cortical actin network are essential for the terminal stages of regulated exocytosis in {beta}-cells.

Address correspondence to: Aniko Varadi (Aniko.Varadi{at}uwe.ac.uk) or Guy A. Rutter (g.a.rutter{at}bris.ac.uk)




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