Myosin Va transports dense core secretory vesicles in pancreatic MIN6 β-cells

Varadi, Aniko; Tsuboi, Takashi; Rutter, Guy A.

Abstract

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 colocalization of myosin Va (MyoVa) with insulin in pancreatic β-cells and show that MyoVa copurifies with insulin in density gradients and with the vesicle marker phogrin-enhanced green fluorescent protein 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 depolarization-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 depolarization-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 β-cells. © 2005 by The American Society for Cell Biology.

• Journal Article Type: Article
• Publication Date: Jun 9, 2005
• Deposit Date: Jan 22, 2010
• Publicly Available Date: Nov 15, 2016
• Journal: Molecular Biology of the Cell
• Electronic ISSN: 1939-4586
• Publisher: American Society for Cell Biology
• Peer Reviewed: Peer Reviewed
• Volume: 16
• Issue: 6
• Pages: 2670-2680
• DOI: https://doi.org/10.1091/mbc.E04-11-1001
• Keywords: Myosin Va transports, dense core secretory vesicles, pancreatic MIN6 β-cells
• Public URL: https://uwe-repository.worktribe.com/output/1050657
• Publisher URL: http://dx.doi.org/10.1091/mbc.E04-11-1001
• Additional Information: Additional Information : This publication was vital for a successful BBSRC New Investigators Award and a Wellcome Trust Project Grant recently awarded to Aniko Varadi.
• Contract Date: Nov 15, 2016